Methods of preservation and flotation for the detection of nematode eggs and coccidian oocysts in faeces of the forest musk deer.

Parasitic infections influence the health of captive forest musk deer (Moschus berezovskii) and affect population increases. Nevertheless, there are few quantitative studies regarding forest musk deer parasites, and there is no common preservation method or flotation solution used for detection of faecal parasites because of the biology of the worms and the host physiological state. The objective of this study was to evaluate preservation and flotation methods for the detection of nematode eggs and coccidian oocysts in faeces of the forest musk deer. The McMaster technique was used to count nematode eggs and coccidian oocysts in 33 samples of faeces. For the nematode eggs, the differences among flotation solutions were significant (P< 0.01), with sodium nitrate being the best flotation solution, and the combination of freezing and sodium nitrate resulted in the greatest number of eggs per gram (EPG = 209.4 ± 67.8). For the coccidian oocysts, the interaction between preservation method and flotation solution was significant (P< 0.01), and the combination of formalin and sodium chloride yielded the greatest number of oocysts per gram (OPG = 1010.7 ± 162.3). The forest musk deer had a high prevalence of parasitic infections, with the parasite load of coccidia (96.4%) significantly greater than that of nematodes (71.9%, P< 0.01). These results confirm that captive forest musk deer suffer from serious parasitic invasions and demonstrate that the novel method described here could be utilized for parasitological diagnosis, detection and prevention in species of Moschidae and Cervidae.

Sensilla on the antennal funiculus of the horse stomach bot fly, Gasterophilus nigricornis.

Gasterophilus nigricornis (Loew) (Diptera: Oestridae) is one of the most damaging obligate parasites of equids in Kalamaili, Xinjiang, China. The main olfactory organs of this stomach bot fly are paired antennae that bear microscopic sensillar structures. The external morphology of the antennal funiculus and sensilla of male G. nigricornis were studied using stereopic microscopy and scanning electron microscopy. A cross-sectional view of the funiculus shows it to be triangular, with an anterodorsal surface, a dorsolateral margin and a posteroventral surface. Almost the entire surface of the funiculus is densely covered with microtrichiae. Small patches lacking these microtrichiae appear as depressions or pits in the surface of the funiculus. Six distinct types of sensilla are recorded, including one trichoid, three basiconic, one auriculate and one clavate sensilla. Trichoid sensilla are the most abundant, followed by the basiconic, auriculate and clavate types in descending order. Only auriculate sensilla are found in pits on the funiculus. Distributions of different sensilla types located on the antennal funiculus are provided. These results are compared with equivalent findings in several other fly species. In addition, the possible functions of the various sensilla types are discussed.

First Report of Wilt on Alfalfa in China Caused by Verticillium nigrescens.

Alfalfa (Medicago sativa Linn.), widely grown throughout the world, is an important perennial forage crop. It is high in protein and digestible fiber and is an excellent source of several vitamins (A, D, E, and K) and minerals for beef cattle, horses, sheep, goats, and even humans (2). Wilt symptoms on alfalfa were observed during a disease survey in Yangling, Shaanxi, China in 2009. Symptoms included discoloration, shortened internode, and plant death. However, the vascular tissue of diseased alfalfa plants did not exhibit discoloration and typical "V" symptoms of Verticillium albo-atrum infection. Eleven fungal isolates were obtained from diseased alfalfa plants in Yangling by a tissue isolation method (1). Isolates were cultured on Czapek Dox Agar (CDA; pH 7.2) slants at 22 ± 1°C in darkness. Colonies on CDA plates were whitish and cream-white when viewed from the underside, later becoming dark gray due to the formation of gray or dark brown chlamydospores in single or in short chains. DNA was extracted from each isolate and the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA) was amplified and sequenced using primers ITS-1F and ITS4. The 11 isolates were divided into five groups based on their in vitro morphological characters. A single isolate from each of the five groups was chosen for ITS sequencing. All five isolates had the same ITS sequence (GenBank Accession No. AB551216). On the basis of the ITS sequence and morphology (4), these isolates were identified as V. nigrescens Pethyhr. (recently renamed as Gibellulopsis nigrescens). Five representative isolates were used to fulfill Koch's postulates. Alfalfa seeds (cv. Cossack) were surface sterilized with 75% ethanol for 5 min, allowed to dry, and planted into cow dung compost that had been autoclaved at 160°C for 2 h. Plants were cultivated under controlled greenhouse conditions at 23 to 25°C with a photoperiod of 14 h. Inoculum was prepared by comminuting 15-day-old cultures and sterile deionized water into a suspension of mycelial fragments and conidia (105 to 106 CFU/ml) in a blender. Seedlings (four-leaf stage) were inoculated by immersing roots in the inoculum suspension for 60 min (3). Each isolate was inoculated onto 30 seedlings, six in each pot; another 30 seedlings were soaked with sterile deionized water for 60 min as a control. After 20 days in the greenhouse, all inoculated plants exhibited wilt symptoms similar to the original wilt symptoms observed on diseased alfalfa plants. In contrast, none of the control plants showed wilt symptoms. The pathogen was reisolated from all diseased plants and confirmed to the original ones. To our knowledge, this is the first report of V. nigrescens infecting M. sativa in China, indicating V. nigrescens as one possible important pathogen of alfalfa. References: (1) O. D. Dhingra and J. B. Sinclair. Basic Plant Pathology Methods. CRC Press, Boca Raton, FL, 1995. (2) D. Jasjeet et al. J. Adv. Sci. Res. 2:50, 2011. (3) H. A. Melouk and C. E. Horner. Phytopathology 64:1267, 1974. (4) R. Zare et al. Nova Hedwigia 85:463, 2007.

Long noncoding RNA LINC00052 suppressed the proliferation, migration and invasion of glioma cells by upregulating KLF6.

OBJECTIVE: Recent studies have discovered a class of dysregulated long noncoding RNAs (lncRNAs) related to carcinogenesis. This study aims to uncover the molecular functions of lncRNA LINC00052 in the tumorigenesis of glioma. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to detect LINC00052 expression in 40 glioma samples and 4 glioma cell lines. Besides, regulatory effects of LINC00052 on the in vitro behaviors of glioma cells were evaluated by the proliferation assay, transwell assay and wound healing assay. Furthermore, the interaction between LINC00052 and kruppel-like factor 6 (KLF6) in mediating the progression of glioma was studied by performing qRT-PCR and Western blot. RESULTS: LINC00052 expression was remarkably downregulated in glioma samples compared with that in adjacent samples. Moreover, cell proliferation, invasion, and migration of glioma were inhibited after overexpression of LINC00052 in vitro. Besides, LINC00052 overexpression upregulated mRNA and protein level of KLF6. Besides, the expression of KLF6 in tumor tissues was positively correlated to the expression of LINC00052. CONCLUSIONS: These results suggested that LINC00052 could repress cell migration, invasion and proliferation in glioma through upregulating KLF6, which may offer a new therapeutic intervention for glioma patients.

[Transformation of common wheat (Triticum aestivum L.) with herbicide-resistant EPSPs gene].

The herbicide-resistant EPSPs (5-enolpyruvylshikimate-3-phosphate synthase) gene was transformed into about 1,000 young spikes and 800 young embryos of wheat variety, Jinghua 1, with gene gun. Thirty-eight and four regenerated plants were obtained respectively screened with glyphosate. All regenerated plants were analysed by PCR and/or Southern blotting. The results indicated that EPSPs gene was integrated stably into the genome of Jinghua 1, and some of the transformants showed fertile. So herbicide-resistant EPSPs gene could be used as selective marker in the transformation of monocotyledon cereal crops, such as wheat.

[Determination of ecdysones in polyploid and monoploid Achyranthes bidentata Bl].

Total ecdysones (ecdysterone and inokosterone) were isolated and identified from Achyranthes bidentata radix of polyploid and monoploid by column chromatography. The hormone contents of monoploid and diploid are about the same, but the content of polyploid is 14 times that of the diploid.