Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples

We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.     

Three‐dimensional CT examination of the forefoot and hindfoot of the hippopotamus and tapir during a semiaquatic walking

Semiaquatic walking has resulted in the evolution of functional and morphological changes in various hoofed mammals, such as hippopotamus and Brazilian tapir. The biomechanics of skilful walking in wetlands or at the bottom of a waterbody involve the medio‐lateral opening and closing of the feet to effectively support and stabilize the body on soft ground and to reduce the water resistance during recovery stroke, respectively. We demonstrate that the opening and closing of the feet in hippopotamus and Brazilian tapir are mediated by the adduction and abduction of the most medial and lateral phalanges from the CT examination. The axial toes, metacarpals and metatarsals do not contribute to changes in the width and shape of the feet, unlike the medial and lateral toes. We suggest that this semiaquatic walking motion is derived from the original terrestrial mode of locomotion, in contrast to the highly functional swimming motion using webs or fins in morphologically modified feet and tail. From the present data we demonstrate that semiaquatic locomotion evolved due to the acquisition of adductor–abductor mobility in the phalanges of the most medial and lateral digits, as shown in hippopotamus and Brazilian tapir.     

Development of two immunochromatographic tests for the serodiagnosis of bovine babesiosis

In this study, we developed two immunochromatographic tests (ICTs), which are nitrocellulose membrane-based immunoassays for the convenient and rapid serodiagnosis of bovine babesiosis caused by Babesia bovis (BoICT) and Babesia bigemina (BiICT). The efficacy of two ICTs was evaluated using 13 positive sera from experimentally infected cattle with B. bovis or B. bigemina. Clear results showed that the BoICT and ELISA detected antibodies in sera collected from 14 to 93 days post-infection, while BiICT and ELISA detected from 13 to 274 days post-infection. In additon, non-infected cattle, Neospora caninum, and Cryptosporidium parvum were negative in two ICTs. To evaluate the field utility of the ICTs, we tested 186 field bovine sera collected from cattle living in Yanbian (China) and Mato Grosso do Sul (Brazil). The results of ICTs were compared to those of classical serodiagnostic methods, enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence assay (IFAT). The overall concordances of BoICT were determined as 92.5 and 90.3% when the results of ELISA and IFAT were set as the reference standards, respectively. In contrast, those of BiICT showed 96.8 and 92.5% relative to the results of standard ELISA and IFAT, respectively. Conventional and rapid diagnosic devices for bovine babesiosis may provide a valuable tool in clinical and field applications.     

Inheritance of gramine content in barley

The inheritance of gramine indole alkaloid was studied in F1 and F2 generations derived from crosses between wild ( Hordeum vulgare subsp. spontaneum) and cultivated ( H. vulgare subsp. vulgare) barley at adult stage in the field. The means of gramine content were very low in F1s and F2s in April and May. The ratio of progeny containing gramine to those without gramine (near zero or undetectable) fit a mono-genic or di-genic model in F2 generation. However, the content of gramine might be controlled by minor genes in the gramine-containing plants. The direction of dominance was toward lower gramine content. The averages of dominance ( h/d) in F1 and F2 ranged from partial to complete. Broad-sense heritabilities were high (0.70 to 0.77). The usefulness of hybrids for breeding aphid resistance and possibility increasing palatability of grazing pasture for sheep and cattle is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Codend selectivity for jack mackerel and whitefin jack and unequal split parameter estimates observed in trouser trawl experiments

Codend selectivity for the jack mackerel Trachurus japonicus and the whitefin jack Kaiwarinus equula were evaluated based on data from trouser trawl experiments carried out in the East China Sea, using a test codend of 60 mm diamond mesh and a control codend made of minnow net with a square mesh of 9 mm bar length. Between-haul variations in parameters and the mean selection curves were tested with the catch data in the SELECT approach, and then the model of between-haul variation in the split parameter with the mean selection curve was chosen as the best fit using Akaike’s information criterion model selection. The 50% retention lengths and the selection ranges were 11.4 and 3.36 cm for jack mackerel and 8.83 and 0.93 cm for whitefin jack, respectively. The selection curve for whitefin jack was sharp, whereas that for jack mackerel was relatively wide. As the estimated split parameters indicated, about 80% of the whitefin jack entered the control codend, but 85 and 90% of the jack mackerel entered the control codend in the second and third hauls, respectively. The inequality in the split parameter is discussed from the viewpoint of the animal’s swimming behavior and water movement based on underwater video observations.     

Binding activity of substituted benzyl derivatives of chloronicotinyl insecticides to housefly‐head membranes,and its relationship to insecticidal activity against the housefly Musca domestica

Variously substituted benzyl derivatives of chloronicotinyl insecticides were synthesized with a wide range of substituents including halogens, NO₂, CN, CF₃ and small alkyl and alkoxy groups at the ortho, meta and para positions, as well as multiple‐substituted benzyl analogues. Their binding activity to the α‐bungarotoxin binding site in housefly (Musca domestica) head membrane preparations was measured. Among the compounds tested, the activity of the meta‐CN derivative was the highest, being 20–100 times higher than those of imidacloprid, acetamiprid and nitenpyram. The synergized insecticidal activity against houseflies was also measured for selected compounds with the metabolic inhibitor, NIA16388 (propargyl propyl phenylphosphonate). For the nitromethylene analogues, including both benzyl and pyridylmethyl analogues, higher binding activity usually resulted in higher insecticidal activity. © 2000 Society of Chemical Industry     

Molecular Structure and Some Physicochemical Properties of High‐Amylose Barley Starches

The molecular structure and some physicochemical properties of starches from two high‐amylose cultivars of barley, high‐amylose Glacier A (HAG‐A) and N (HAG‐N), were examined and compared with those of a normal cultivar, Normal Glacier (NG). The true amylose contents of HAG‐A, HAG‐N, and NG were 41.0, 33.4, and 23.0%, respectively. Iodine affinities before and after defatting of starch, and thermograms of differential scanning calorimetry, indicated that HAG‐A and HAG‐N starches had a higher proportion of amylose‐lipid complex than did NG starch. The amylopectins from HAG‐A and HAG‐N were similar to NG amylopectin in average chain length (18–19), β‐amylolysis limit (β‐AL 56–57%), number‐average degrees of polymerization (DP_n 6,000–7,500) and chain length distribution. Very long chains (1–2%) were found in amylopectins from all cultivars. HAG‐A amylopectin had a larger amount of phosphorus (214 ppm) than the others. The amyloses from HAG‐A and HAG‐N resembled NG amylose in DP_n (950–1,080) and β‐AL (70–74%). However, HAG‐A and HAG‐N had a larger number of chains per molecule (NC 2.4–2.7) than NG amylose (1.8) and contained the branched amylose with a higher NC (9.5–10.6) than that of NG amylose (5.8), although molar fractions of the branched amylose (15–20%) were similar.     

Expressed sequence tag analysis of phyllosomas and hemocytes of Japanese spiny lobster Panulirus japonicus

Two complementary DNA (cDNA) libraries were constructed from phyllosomas and hemocytes of adult Japanese spiny lobster Panulirus japonicus and a total of 2,673 expressed sequence tags (ESTs) were obtained. After assembly and clustering, 450 and 458 unique sequences were found from the phyllosoma and hemocyte cDNA libraries, respectively. Of these, 114 and 220 ESTs showed significant homologies with known genes in the National Centre for Biotechnology Information (NCBI) database. The remaining sequences were of unknown function. Immune-related genes found in this study include lectin, proteinase inhibitor, prophenoloxidase, heat-shock protein, antimicrobial peptide, and a few putative defense-related proteins.     

Do Horses Prefer Certain Substrates for Rolling in Grazing Pasture?

We assessed whether rolling damage by grazing horses could be reduced by constructing areas assigned for rolling. A group of horses were enclosed in a paddock with and without rolling areas made of dry soil, sand, and straw. Their behavior was recorded for 1 week in the paddock without any treatment (control paddock). Then the horses were moved to another paddock with the rolling areas (rolling paddock). After a 3-week familiarization period, horses were observed for 1 week. In the rolling paddock, the frequency and time spent rolling were significantly greater in rolling areas than in nonrolling areas. Horses significantly preferred the soil rolling area than sand and straw (P < .05). Although rolling was considered the most relevant body care behavioral element, the effects of the substrate in rolling areas on other body care behavioral activities, such as mutual and self-grooming, also were investigated. The frequency and duration of mutual grooming and the duration of self-grooming decreased significantly in the rolling paddock compared with the control paddock (P < .05). Hence, offering a rolling area encourages horses to roll in these areas and keeps the pasture in good condition; therefore, grazing time can be increased, with less reliance on supplementary feed.