Occurrence of Theileria and Babesia species in water buffalo (Bubalus babalis, Linnaeus, 1758) in the Hubei province, South China

The presence and prevalence of tick-borne haemoparasites in water buffalo from the Hubei province, south China was investigated using the reverse line blot (RLB) hybridization assay and phylogenetic analysis of the parasite 18S rRNA gene. Theileria buffeli (19.1%) was the most frequently found species in all of the locations, followed by Babesia orientalis (8.9%), Babesia bovis (1.0%) and Babesia bigemina (0.7%). Only 12 (3.9%) of the samples had mixed infections. Eleven samples with single infections were selected for further characterization using 18S rRNA gene sequence analysis. Phylogenetic analysis showed that the eight T. buffeli 18S rRNA gene sequences obtained grouped into four clusters, of which three grouped with the known T. buffeli types B and D. The remaining five grouped separately from the previously describe T. buffeli types, constituting new T. buffeli types. The two B. bigemina 18S rRNA gene sequences obtained grouped closely with B. bigemina Kunming; this serves as the first report of B. bigemina in the Hubei province. The B. orientalis Daye 18S rRNA gene sequence obtained grouped closely with the previously reported B. orientalis Wuhan strain and with Babesia sp. Kashi 1 and Kashi 2.     

Comparison of sorption and diffusion by pyridate and its polar metabolite in isolated cuticular wax of Chenopodium album and Hordeum vulgare

Sorption and diffusion of the herbicide pyridate and its metabolite CL9673 were measured in reconstituted cuticular waxes isolated from Chenopodium album L. and Hordeum vulgare L. (cultivar Igri) leaves. The compounds have the same basic chemical structure, except that pyridate is characterized by a C8-alkyl chain bound via a thioester to the ionizable hydroxyl group of CL9673. Sorption of the weak acid CL9673 from aqueous solutions into cuticular waxes was pH-dependent, and the apparent wax/water partition coefficients decreased with increasing pH. Wax/water partition coefficients of pyridate were not dependent on pH, and they were about 4 orders of magnitude higher as compared to the nondissociated species of CL9673. Diffusion coefficients measured in reconstituted cuticular wax for CL9673 fitted established models predicting diffusion coefficients in relation to molar volumes. However, this was not the case with pyridate, which was characterized by a self-accelerating effect leading to diffusion coefficients, which were up to 2 orders of magnitude higher than predicted from the molar volume. This is a remarkable result since pyridate represents a compound combining the properties of an active ingredient and of a plasticizer.     

Interactions of calcium ions with weakly acidic active ingredients slow cuticular penetration: a case study with glyphosate

Potassium and calcium salts of glyphosate were obtained by titrating glyphosate acid with the respective bases to pH 4.0, and rates of penetration of these salts across isolated astomatous cuticular membranes (CMs) were measured at 20 degrees C and 70, 80, 90, and 100% humidity. K-glyphosate exhibited first-order penetration kinetics, and rate constants (k) increased with increasing humidity. Ca-glyphosate penetrated only when the humidity above the salt residue was 100%. At 90% humidity and below, Ca-glyphosate formed a solid residue on the CMs and penetration was not measurable. With Ca-glyphosate, the k value at 100% humidity decreased with time and the initial rates were lower than for K-glyphosate by a factor of 3.68. After equimolar concentrations of ammonium oxalate were added to Ca-glyphosate, high penetration rates close to those measured with K-glyphosate were measured at all humidities. Adding ammonium sulfate or potassium carbonate also increased rates between 70 and 100% humidity, but they were not as high as with ammonium oxalate. The data indicate that at pH 4.0 one Ca2+ ion is bound to two glyphosate anions. This salt has its deliquescence point near 100% humidity. Therefore, it is a solid at lower humidity and does not penetrate. Its molecular weight is 1.82 times larger than that of K-glyphosate, and this greatly slows down rates of penetration, even at 100% humidity. The additives tested have low solubility products and form insoluble precipitates with Ca2+ ions, but only ammonium oxalate binds Ca2+ quantitatively. The resulting ammonium salt of glyphosate penetrates at 70-100% humidity and at rates comparable to K-glyphosate. The results contribute to a better understanding of the hard water antagonism observed with glyphosate. It is argued that other pesticides and hormones with carboxyl functions are likely to respond to Ca2+ ions in a similar fashion. In all of these cases, ammonium oxalate is expected to overcome hard water antagonism.     

Piroplasm parasites of white rhinoceroses (Ceratotherium simum) in the Kruger National Park, and their relation to anaemia

As part of a larger survey to map the geographical distribution of Babesia and Theileria parasites in the southern African rhinoceros population, white rhinoceroses were sampled during routine immobilisations in the Kruger National Park. Polymerase chain reaction (PCR) and reverse line blot (RLB) hybridisation assays were used to screen for the presence of piroplasms and complete blood counts were used to assess associated changes in clinical parameters. Of the 195 rhinoceroses sampled, 71 (36.4 %) tested positive for the presence of Theileria bicornis, with no significant change in the haematological parameters measured, while 18 (9.2 %) tested positive for Theileria equi. None of the rhinoceroses sampled tested positive for Babesia bicornis, a parasite associated with mortalities in black rhinoceroses.     

Identification of Theileria parva and Theileria sp. (buffalo) 18S rRNA gene sequence variants in the African Buffalo (Syncerus caffer) in southern Africa

Theileria parva is the causative agent of Corridor disease in cattle in South Africa. The African buffalo (Syncerus caffer) is the reservoir host, and, as these animals are important for eco-tourism in South Africa, it is compulsory to test and certify them disease free prior to translocation. A T. parva-specific real-time polymerase chain reaction (PCR) test based on the small subunit ribosomal RNA (18S rRNA) gene is one of the tests used for the diagnosis of the parasite in buffalo and cattle in South Africa. However, because of the high similarity between the 18S rRNA gene sequences of T. parva and Theileria sp. (buffalo), the latter is also amplified by the real-time PCR primers, although it is not detected by the T. parva-specific hybridization probes. Preliminary sequencing studies have revealed a small number of sequence differences within the 18S rRNA gene in both species but the extent of this sequence variation is unknown. The aim of the current study was to sequence the 18S rRNA genes of T. parva and Theileria sp. (buffalo), and to determine whether all identified genotypes can be correctly detected by the real-time PCR assay. The reverse line blot (RLB) hybridization assay was used to identify T. parva and Theileria sp. (buffalo) positive samples from buffalo blood samples originating from the Kruger National Park, Hluhluwe-iMfolozi Park, the Greater Limpopo Transfrontier Park, and a private game ranch in the Hoedspruit area. T. parva and Theileria sp. (buffalo) were identified in 42% and 28%, respectively, of 252 samples, mainly as mixed infections. The full-length 18S rRNA gene of selected samples was amplified, cloned and sequenced. From a total of 20 sequences obtained, 10 grouped with previously published T. parva sequences from GenBank while 10 sequences grouped with a previously published Theileria sp. (buffalo) sequence. All these formed a monophyletic group with known pathogenic Theileria species. Our phylogenetic analyses confirm the distinction between Theileria sp. (buffalo) and T. parva and indicate the existence of a single group of T. parva and two Theileria sp. (buffalo) 18S rRNA gene variants in the African buffalo. Despite the observed variation in the full-length parasite 18S rRNA gene sequences, the area in the V4 hypervariable region where the RLB and real-time PCR hybridization probes were developed was relatively conserved. The T. parva specific real-time PCR assay was able to successfully detect all T. parva variants and, although amplicons were obtained from Theileria sp. (buffalo) DNA, none of the Theileria sp. (buffalo) 18S rRNA sequence variants were detected by the T. parva-specific hybridization probes.     

Physiological and behavioural responses of grazing dairy cows to an acute metabolic challenge

Due to seasonal changes in the quality and quantity of herbage, the nutrient supply to grazing dairy cows is not always sufficient, which may increase their metabolic load. To investigate the temporal pattern of behavioural changes in relation to concomitant metabolic alterations, we subjected 15 multiparous early lactating Holstein dairy cows (24 (SD 7.4) days in milk) to a short‐term metabolic challenge, which we provoked by abruptly withdrawing concentrate for 1 week. Cows grazed full‐time and were supplemented with concentrate in experimental week (EW) 1 and EW 3, whereas concentrate was withdrawn in EW 2. We analysed milk and blood samples to characterise the metabolic changes and found that the total yield of milk and protein decreased (p < 0.05) and fat yield, fat‐to‐protein ratio and acetone content increased (p < 0.05) from EW 1 to EW 2. Plasma glucose and insulin concentrations were lower (p < 0.05), and concentrations of nonesterified fatty acids and beta‐hydroxybutyrate were higher (p < 0.05) in EW 2 compared with EW 1. Apart from ingestive and rumination behaviour and activity, we also monitored the use of an automated brush on pasture. While time spent eating and ruminating increased (p < 0.05) in EW 2 compared with EW 1, time spent idling decreased (p < 0.05). Concomitantly, while time standing and moving increased (p < 0.05) from EW 1 to EW 2, walking time decreased (p < 0.05). The daily proportion of cows using the automated brush decreased (p < 0.05) in EW 2 compared with EW 1, as did the duration of brushing per day. In conclusion, grazing cows experiencing a metabolic challenge try to compensate for the nutrient deficiency by increasing eating time, a behavioural element important for short‐term survival. Due to the strong impact of weather conditions, we cannot currently recommend observation of outdoor brushing activity to address short‐term alterations in the metabolic state of grazing cows.     

PhenoFly Planning Tool: flight planning for high-resolution optical remote sensing with unmanned areal systems

Background

Driven by a huge improvement in automation, unmanned areal systems (UAS) are increasingly used for field observations and high-throughput phenotyping. Today, the bottleneck does not lie in the ability to fly a drone anymore, but rather in the appropriate flight planning to capture images with sufficient quality. Proper flight preparation for photography with digital frame cameras should include relevant concepts such as view, sharpness and exposure calculations. Additionally, if mapping areas with UASs, one has to consider concepts related to ground control points (GCPs), viewing geometry and way-point flights. Unfortunately, non of the available flight planning tools covers all these aspects.

Results

We give an overview of concepts related to flight preparation, present the newly developed open source software PhenoFly Planning Tool, and evaluate other recent flight planning tools. We find that current flight planning and mapping tools strongly focus on vendor-specific solutions and mostly ignore basic photographic properties—our comparison shows, for example, that only two out of thirteen evaluated tools consider motion blur restrictions, and none of them depth of field limits. In contrast, PhenoFly Planning Tool enhances recent sophisticated UAS and autopilot systems with an optical remote sensing workflow that respects photographic concepts. The tool can assist in selecting the right equipment for your needs, experimenting with different flight settings to test the performance of the resulting imagery, preparing the field and GCP setup, and generating a flight path that can be exported as waypoints to be uploaded to an UAS.

Conclusion

By considering the introduced concepts, uncertainty in UAS-based remote sensing and high-throughput phenotyping may be considerably reduced. The presented software PhenoFly Planning Tool (https://shiny.usys.ethz.ch/PhenoFlyPlanningTool) helps users to comprehend and apply these concepts.

     

Spatial patterns and temporal changes of heavy metal distributions in river sediments in a region with multiple pollution sources

Purpose

The main purpose of this study was to evaluate temporal and regional variability of contamination by heavy metals (HMs) in river sediments using their enrichment factors (EFs) and benchmarking according to sediment quality guidelines (SQGs). The Zlin region in the Czech Republic (Morava and Drevnice River basins) represents a model area where several regionally specific ecological risk assessment studies have recently been conducted with a focus on organic pollution, eco-toxicity, geological, and geochemical characteristics.

Materials and methods

Four consecutive sediment sampling campaigns were undertaken in spring and autumn 2005–2006. Aqua-regia leachable content of Cd, Co, Cr, Cu, Ni, Pb, Sb, V, and Zn in surface sediments from 14 sites was analyzed using ICP-MS, and Hg content was analyzed using AMA-254 analyzer. EFs were calculated to identify the human impact on pollution in the area. Comparisons to SGQs were conducted to identify the areas and HMs of greatest risk.

Results and discussion

Calculation of EFs contributed to the effective clustering of HMs. Median EFs of Co, Ni, and V ranged from 0.9 to 1.4 at all sites indicating concentrations very close to natural geological background levels. There was greater enrichment at locally polluted sites, the highest in the cases of Cd, Sb, Hg, and Cr. Widespread influence of diffuse HM sources (traffic, agriculture, and urban wastes) was apparent from elevated concentrations of Pb, Cu, and Zn at all sites. EF values also helped to identify the greatest temporal changes and shifts in HMs contamination between adjacent sites caused by 50-year recurrence interval floods in early spring 2006. The impact was most apparent in downstream sites; namely directly below the confluence of the two major rivers.

Conclusions

The overall contamination of HMs in the region was classified as low-to-moderate with significantly contaminated sub-areas. The study showed relatively stable spatial distributions of HMs, indicating potential sources of pollution. Cu was identified as the HM of greatest risk. The study emphasizes the necessity of considering both environmental circumstances and background HM occurrence to prevent misinterpretation of the pollution situation. The use of EFs which include grain size proxy normalization and HM background levels, along with the comparison of the detected concentrations to SQGs, proved an efficient way to identify hazardous contamination from anthropogenic sources.     

USE OF CONTRAST‐ENHANCED COMPUTED TOMOGRAPHY TO STUDY THE CRANIAL MIGRATION OF A LUMBOSACRAL INJECTATE IN CADAVER DOGS

Volumes used in lumbosacral epidural injections for anesthesia have remained unchanged since the 1960s. The goals of this cross‐sectional observational study were to characterize the three‐dimensional spread of a lumbosacral epidural injection, as well as confirm that the commonly used volume of 0.2 ml/kg injected into the lumbosacral epidural space reaches the thoracolumbar (TL) junction in the majority (≥80%) of dogs. Ten clinically normal, adult, nonpregnant, mixed‐breed dogs were obtained within five minutes of euthanasia and 0.2 ml/kg of radiopaque contrast medium was injected into the lumbosacral epidural space. A computed tomography scan of the TL spine was performed immediately following the injection. Migration of contrast reached the TL junction in 8 of 10 (80%) dogs. Contrast was well visualized in all epidural planes with contrast travelling predominantly in the dorsal epidural space in 7 of 10 (70%) dogs. There was no significant difference in the weight of dogs where the epidural injectate reached the TL junction and those where it did not (P = 0.16), or in the weight of dogs where the cranial‐most point of the contrast column was in the dorsal versus the ventral epidural space (P = 0.32). This preliminary study supports the use of computed tomography to characterize injectate distribution in the canine thoracolumbar epidural space and provides evidence that a 0.2‐ml/kg volume is likely to reache the TL junction in most dogs. Further studies are needed in live dogs to determine if variables affecting human epidural injectate doses have similar effects in the dog.