Case of suspected theobromine poisoning in dairy cattle

High-yielding dairy cows collapsed and died displaying signs of a disturbed central nervous system (muscle tremor, convulsion) and a considerably reduced body condition score. An intense diagnostic screening did not allow to confirm a definite diagnosis. Therefore, further analyses including an evaluation of feeds and feeding were initiated. The herd was fed a total mixed ration (TMR) based on corn and grass silage of moderate nutritive value supplemented with various amounts of chocolate chips. In retrospect, the amount of chocolate chips added to the respective TMR batches could not be quantified. These chips were purchased from a company producing bakery and chocolate products for human consumption and added to the silage and therefore to the TMR in order to increase the energy intake of the animals. Because the TMR, which was fed during the time of the incidence, was no longer available, a sample of a later batch of silage was examined. The amount of chocolate chips was quantified (0.44% per fresh matter), and a theobromine analysis was performed in the chocolate chips of the available batch (69.7 mg/100 g fresh matter). Because of the possible link between chocolate intake and observed signs, an immediate cessation of using the chocolate chips was recommended in addition to an optimisation of the TMR, that is an increase of the energy density. Even though the theobromine intake during the time of the incidence is unknown and information about toxicity of theobromine in ruminants is limited, we suspect that the feeding of chocolate in this case caused all signs including the sudden death of the cows. Further reasons are that no differential diagnoses were established and the problems at the farm stopped after removing the chocolate from the TMR.     

Comparison of core needle biopsy and fine-needle aspiration of enlarged peripheral lymph nodes for antemortem diagnosis of enzootic bovine lymphosarcoma in cattle

OBJECTIVE: To determine whether antemortem core needle biopsy and fine-needle aspiration of enlarged peripheral lymph nodes could be used to distinguish between inflammation and lymphosarcoma in cattle. DESIGN: Prospective study. ANIMALS: 25 cattle with enlarged peripheral lymph nodes. PROCEDURES: Antemortem biopsies of the selected lymph nodes were performed with an 18-gauge, 12-cm core needle biopsy instrument. Fine-needle aspirates were performed with a 20-gauge, 4-cm needle. Specimens were analyzed by pathologists who were unaware of clinical findings and final necropsy findings, and specimens were categorized as reactive, neoplastic, or nondiagnostic for comparison with necropsy results. RESULTS: Sensitivity and specificity of core needle biopsy ranged from 38% to 67% and from 80% to 25%, respectively. Sensitivity of fine-needle aspiration ranged from 41% to 53%, and specificity was 100%. Predictive values for positive test results ranged from 77% to 89% for core needle biopsy and were 100% for fine-needle aspiration. Predictive values for negative test results were low for both core needle biopsy and fine-needle aspiration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that core needle biopsy and fine-needle aspiration can aid in the antemortem diagnosis of bovine enzootic lymphosarcoma. Results of fine-needle aspiration of enlarged peripheral lymph nodes were more specific and more predictive for a positive test result than were results of core needle biopsy.     

Indigenous protected areas in Sea Country: Indigenous‐driven collaborative marine protected areas in Australia

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Leaching of carbon and nitrogen from a sandy soil substrate: A comparison between suction plates and ion exchange resins

To determine boundary effects on leaching, we investigated (1) how filter materials affect the concentrations of dissolved organic carbon (DOC) and nitrate (NO₃‐N) in soil percolates and (2) whether ion exchange resins and suction plates are equally suited to capture NO₃‐N. DOC leaching was higher with PE suction plates and plate material did not affect NO₃‐N leachate concentrations. Cumulative NO₃‐N leaching was similar for glass suction plates and ion exchange resins.     

Binding and bioactivity of insulin in primary cultures of carp (Cyprinus carpio) hepatocytes

Isolated carp hepatocytes cultured in serum-free, chemically defined medium were used to investigate within the same cell preparation characteristics of the binding of insulin as well as effects of insulin on cellular metabolism. The binding of human [¹²⁵I]-insulin to carp hepatocytes was studied in kinetic, saturation and displacement experiments. A dependency of insulin binding on the collagenase used for cell isolation was demonstrated. Insulin binding decreased during the first 12h of culture but remained constant during the following 12h. The kinetic experiments revealed that [¹²⁵I]-insulin binding reached a steady state within 20–30 min of incubation. The mathematical analysis of the saturation experiments demonstrated the existence of two populations of binding sites, one with high affinity (K_d1 = 5.5 pM) and low capacity (B_max1 = 0.14 fmol/mg protein or 77 binding sites/cell) and one with low affinity (K_d2 = 2.4 nM) and high capacity (B_max2 = 17.6 fmol/mg protein or 9623 binding sites/cell). In competition experiments, 312 pM [¹²⁵I]-insulin was displaced by cold insulin, IGF-I and IGF-II with IC₅₀ values of 2.2, 7.9 and 20.3 nM, respectively. Glucagon was without effect. Binding of insulin to carp hepatocytes resulted in a significant reduction of glucose release and a significant increase of protein synthesis as of de novo fatty acid synthesis. dedicated to Prof. Dr. W. Hanke on the occasion of his 65^th birthday.     

Reproductive performance of cultured brown trout (Salmo trutta L.) in Chile

Data on reproductive traits in a cultured population of brown trout (Salmo trutta L.) in southern Chile were collected over three consecutive reproductive seasons: 1999, 2000 and 2001, corresponding to the first (at 3 years), second and third spawnings in females. Data were collected from individual females (n=238, 273 and 169 respectively). The reproductive season lasted mainly from June to August. The peak (July) tended to increase with each season (55.46%, 62.27% and 80.81% of total spawning fish respectively). Female body weight (470.8±102.5, 735.0±150 and 1263.9±263.4 g), total fecundity (1182±344, 1904±595 and 2744±605) and egg diameter (4.64±0.11, 4.67±0.27 and 5.24±0.12 mm) increased significantly (P<0.01) over successive reproductive seasons, particularly between the second and third spawnings. Relative fecundity, on the other hand, decreased significantly with each season (3577±471, 2591±900 and 2181±360). Following analysis of the variables over the three seasons, the correlation pattern of female body weight with total fecundity (r=0.91, n=458; P<0.001) and relative fecundity (r=?034, n=451; P<0.001) proved similar to that described in other salmonid females. Fertilization rate and survival to the eyed egg stage were also positively correlated (r=0.73, n=453; P<0.001). The systematically high values obtained for these latter variables over the seasons evaluated (consistently above 90%) are clearly greater than those we registered in other species of salmonids bred in Chile under similar conditions and suggest highly efficient biological variables that determine the gamete fertilization of this species. The brown trout is, therefore, an interesting potential aquaculture resource in Chile.     

Influence of nutrition on the lipid quality of pond fish: common carp (Cyprinus carpio) and tench (Tinca tinca)

Like marine fish freshwater fish are an important source of essential fatty acids for human nutrition. However, the fatty acid composition of pond fish can vary considerably and strongly depends on that of the ingested food. Investigations on the fatty acid composition of common carp (Cyprinus carpio) and tench (Tinca tinca) have shown that different methods of rearing and feeding cause substantial variations in the proportions of the n-6 and n-3 polyunsaturated fatty acids of these fish species. Carp reared on the basis of natural food in ponds exhibit high contents of n-6 as well as n-3 fatty acids in their muscle triacylglycerols. On the other hand carp fed supplementary wheat in ponds resulted in somewhat lower levels of these essential fatty acids. High amounts of n-3 fatty acids can be found in carp fed high-energy diets containing high levels of fish oil. Analogous results were obtained in experiments with tench reared under different nutritional conditions. While rearing on the basis of only natural food in ponds as well as feeding supplementary wheat yielded in similar levels of n-3 and n-6 polyunsaturated fatty acids, higher contents of n-3 fatty acids were recorded in tench fed pellets. High levels of n-3 polyunsaturated fatty acids in foodstuffs have positive effects on human health. Experiments with different cultured fish species proved that the fatty acid composition of the edible parts can be influenced by the diet. Therefore, a finishing diet with a suitable fatty acid profile can be used to improve the nutritional quality of fish products of farmed origin.     

Outbreak of Philophthalmus gralli in four greater rheas (Rhea americana)

Using slit‐lamp biomicroscopy, conjunctival biopsy, and morphological identification, a flock of four Greater rheas (Rhea americana) in Arizona were diagnosed with conjunctivitis secondary to Philophthalmus gralli (P. gralli) infection. Aquatic snails from the exhibit’s water source were identified as Melanoides tuberculatus, a known vector for P. gralli. Comparison of partial sequences of DNA regions from P. gralli adults removed from the rheas and metacercariae from the aquatic snails demonstrated a 100% match, confirming the source of infection. The flock was divided into two treatment groups: the most severely affected rheas received both manual removal of trematodes and praziquantel 1% ointment OU q12 h and the least severely affected rheas were only given praziquantel 1% ointment OU q12 h. The rheas were permanently relocated away from the infected water source and aquatic snails. Initial resolution was seen at 17 weeks in the most severely affected rhea, which had 675 adult P. gralli removed and topical praziquantel. The two rheas that only received topical praziquantel showed resolution within 3 and 15 weeks. Current recommendations for treating P. gralli include: manual removal of trematodes, topical praziquantel 1% ointment, and relocation away from infected water sources and aquatic snails.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Bioindication and modelling of atmospheric deposition in forests enable exposure and effect monitoring at high spatial density across scales

Key message

Moss surveys provide spatially dense data on environmental concentrations of heavy metals and nitrogen which, together with other biomonitoring and modelling data, can be used for indicating deposition to terrestrial ecosystems and related effects across time and areas of different spatial extension.

Context

For enhancing the spatial resolution of measuring and mapping atmospheric deposition by technical devices and by modelling, moss is used complementarily as bio-monitor.

Aims

This paper investigated whether nitrogen and heavy metal concentrations derived by biomonitoring of atmospheric deposition are statistically meaningful in terms of compliance with minimum sample size across several spatial levels (objective 1), whether this is also true in terms of geostatistical criteria such as spatial auto-correlation and, by this, estimated values for unsampled locations (objective 2) and whether moss indicates atmospheric deposition in a similar way as modelled deposition, tree foliage and natural surface soil at the European and country level, and whether they indicate site-specific variance due to canopy drip (objective 3).

Methods

Data from modelling and biomonitoring atmospheric deposition were statistically analysed by means of minimum sample size calculation, by geostatistics as well as by bivariate correlation analyses and by multivariate correlation analyses using the Classification and Regression Tree approach and the Random Forests method.

Results

It was found that the compliance of measurements with the minimum sample size varies by spatial scale and element measured. For unsampled locations, estimation could be derived. Statistically significant correlations between concentrations of heavy metals and nitrogen in moss and modelled atmospheric deposition, and concentrations in leaves, needles and soil were found. Significant influence of canopy drip on nitrogen concentration in moss was proven.

Conclusion

Moss surveys should complement modelled atmospheric deposition data as well as other biomonitoring approaches and offer a great potential for various terrestrial monitoring programmes dealing with exposure and effects.