Search

Human echinococcosis is present worldwide but it is in China that disease prevalence is the highest. In western China, especially in the Tibetan Plateau, the burden of echinococcosis is the most important. Dogs are a major definitive host of Echinococcus and monitoring the presence of Echinococcus worms in dogs is therefore essential to efficiently control the disease. Detection kits based on three different technologies including sandwich ELISA, (indirect) ELISA, and gold immunodiffusion, are currently marketed and used in China. The objective of this work was to assess the efficacy of these kits, in particular with respect to sensitivity and specificity. Four fecal antigen detection kits for canine infection reflecting the three technologies were obtained from companies and tested in parallel on 220 fecal samples. The results indicate that the performance is lower than expected, in particular in terms of sensitivity. The best results were obtained with the sandwich ELISA technology. The gold immunofiltration yielded the poorest results. In all cases, further development is needed to improve the performance of these kits which are key components for the control of echinococcosis., Author summary Although present worldwide, human echinococcosis is at its highest prevalence in western China and particularly in the Tibetan Plateau. Controlling echinococcosis is a national priority and routine monitoring must be established. Dogs are the main infection source in China and surveying Echinococcus worms in dogs is therefore a research priority. Commercial detection kits are currently in use in China to monitor the presence of Echinococcus in dogs. The kits are based on three different technologies including sandwich ELISA with two monoclonal antibodies, (indirect) ELISA, and gold immunodiffusion. National survey programmes are essential for the control of echinococcosis and it is thus very important to assess the efficacy of these kits for the programmes. This work was therefore undertaken to assess efficacy of the kits, in particular with respect to sensitivity and specificity. Four fecal antigen detection kits for canine infection reflecting the three technologies were obtained from companies and tested in parallel on 220 fecal samples. The performance was lower than expected, in particular for their sensitivity which ranged between 51.5% and 83.9% with only two samples displaying a worm burden lower than 100. Three out of four kits showed non-specific cross-reactions with other parasites. The best results were obtained with the sandwich ELISA technology, whereas gold immunofiltration yielded the poorest results. However, in all cases, further development is strongly needed to improve the performance of these kits which are key components for the control of echinococcosis.

Loropetalum subcordatum is a national key protected species endemic to China. Despite its ecological importance and horticultural value, the whole chloroplast (cp) genome of it has not been reported yet. Here, using the paired-end shotgun sequencing technology, we determined the first nucleotide sequence of L. subcordatum cp genome. Our results showed the complete cp genome of L. subcordatum was 158,706 bp long and displayed a typical quadripartite structure which consisted of a pair of inverted repeats (IRs) with a length of 25,998 bp, separating by two single-copy regions (LSC, 88,216 bp and SSC, 18,494 bp). Besides, the cp genome of L. subcordatum contained 131 genes, including 86 protein coding genes, 37 transfer RNAs and 8 ribosomal RNAs. Furthermore, we identified a total of 63 microsatellites (SSRs) within the L. subcordatum cp genome. These newly developed genomic resources will greatly helpful to the further conservation genetic studies of this endangered tree.

Atractylodes macrocephala is one of the most commonly used herbs in China, which is famous for its high medicinal value. In this study, we analyzed and characterized the complete plastome sequence of A. macrocephala. Sequence analysis indicated that the entire genome is 153,265 bp in length, consisting of a large single-copy (LSC, 84,311 bp) and a small single-copy (SSC, 18,674 bp) region separated by a pair of inverted repeat (IR) regions of 25,140 bp for each. The genome contains 107 unique genes, including 80 different protein-coding genes, 23 tRNA genes, and 4 rRNA genes. The overall GC content of the genome is 37.7%. The phylogenetic analysis revealed a monophyletic Atractylodes and Cardueae. This research reports the complete plastome genome of Atractylodes macrocephala, which provides a better understanding of this important herb.

Background Echinococcoses, caused by metacestodes of Echinococcus granulosus (cystic echinococcosis; CE) and E. multilocularis (alveolar echinococcosis; AE), represent major emerging parasitic diseases. These enzootic helminthiases invoke significant public health concerns and social burdens in endemic areas. The diseases are prevalent in the Qinghai-Tibetan Plateau, China, while community-based epidemiological studies have been scarcely reported. We surveyed echinococcosis patients in the southeastern Qinghai Province, China, to better understand the concurrent epidemiological situation in this area. Methods During July and August of 2013 and 2014, we screened echinococcosis patients at Yushu and Golog Prefectures, Qinghai Province, China, in a diagnostic campaign. A total of 2856 people (male:female ratio, 1:1.12; mean age, 34.6 years; age range, 6–88 years) were ultrasonographically examined for the presence of hepatic echinococcal cysts. We also collected serum samples from patients and analyzed antibody reactivity against recombinant forms of diverse E. granulosus antigen Bs (rEgAgB1-5) by enzyme-linked immunosorbent assay. Results We detected 134 patients whose imaging scans were compatible with CE (115 cases) and AE (20 patients). One patient might have been infected with both CE and AE. The overall incidence was 4.7% (CE, 4.0%; AE, 0.7%). A large proportion (67.5%) of CE patients was diagnosed at active and transitional CE1-CE3 stages in their late 30s. The AE cases were generally detected at advanced stage in patients at early 20s (60%). Analysis of the receiver operating characteristic curve and Youden’s index indicated that rEgAgB2 was the most promising biomarker, followed by rEgAgB3 and rEgAgB1. Overall, sensitivity and specificity of rEgAgB1-3 were 84.5–92.7% and 91.9–94.6%, respectively. rEgAgB4 and 5 showed low sensitivity with high cross-reactivity. Conclusions Our results strongly suggest that disability-adjusted life years related to echinococcoses in Qinghai-Tibetan areas might be more serious than previously considered. Control and prevention strategy against CE and AE are highly required in these areas. In addition to ultrasonography, serological tests might provide supportive data. However, serological data should be carefully interpreted for differential diagnosis, especially in areas where both CE and AE are co-endemic. Electronic supplementary material The online version of this article (10.1186/s13071-019-3569-6) contains supplementary material, which is available to authorized users.

Cystic echinococcosis (CE) is a common, chronic, and endemic zoonotic disease usually localized in a single organ; multivisceral cases are rare, especially outside the liver or lung. Here, we describe an unusual case of a 43-year-old Tibetan man with echinococcosis of the infratemporal fossa, heart, liver, pancreas, abdomen, and pelvic cavity. He only presented with diminished vision of the left eye, especially when chewing. Computed tomography and magnetic resonance imaging revealed multivisceral CE. The patient underwent surgery for the excision of a cyst in the infratemporal fossa, as well as chemotherapy, and the diagnosis was confirmed by histopathological examination. The diagnosis, clinical features, treatment, and follow-up in this case are discussed. In areas with high echinococcosis prevalence, examination by full imaging is necessary for an accurate diagnosis, especially in cases of atypical localization. Chemotherapy for treatment, as well as prophylaxis against recurrence, can be effective when surgery is not possible.

Additional file 2: Table S1. Gene-specific oligonucleotide primers used.

Alveolar echinococcosis (AE), caused by the Echinococcus multilocularis metacestode, represents one of the most frequently fatal zoonoses. Early diagnosis significantly reduces morbidity and mortality associated with AE. Diagnosis of AE largely depends on a combination of imaging and serological tests due to its minimal clinical manifestations. Several antigens derived from the whole worm and protoscolex have been targeted for AE serodiagnosis, while the antigenic properties of E. multilocularis hydatid fluid (EmHF) are unclear. We observed two AE-specific 6- and 8-kDa antigen proteoforms through an immunoproteome array of the EmHF. We identified these proteins as representing an E. multilocularis antigen B3 (EmAgB3) isoform, and the proteins were shown to be encoded by the same gene. We cloned the gene and expressed the recombinant EmAgB3 protein (rEmAgB3) in Escherichia coli . rEmAgB3 exhibited sensitivity of 90.9% (80/88 cases) and specificity of 98.5% (597/606 samples) by immunoblotting. The positive and negative predictive values were 89.9% and 98.6%, respectively. The protein did not show antibody responses to 33 AE sera collected during posttreatment follow-up monitoring. Mouse sera experimentally infected with AE protoscoleces began to demonstrate specific antibody responses to native and recombinant EmAgB3 6 months after infection. At that stage, fully mature metacestode vesicles that harbored the brood capsule, primary cell, and protoscolex were observed within an AE mass(es). The response declined along with worm degeneration. Our results demonstrate that the immune responses to this EmAgB3 isoform were highly correlated with worm viability accompanied with AE progression. rEmAgB3 is a promising biomarker for serological assessment of AE patients.

A method was developed for simultaneous determination of chloramphenicol (CAP), thiamphenicol (TAP), and florfenicol (FF) residues in animal tissues using gas chromatography/mass spectrometry (GC/MS) with chemical ionization source in negative mode. The homogenized samples were extracted with ethyl acetate and the extracts were partitioned with n -hexane to remove lipids. Further cleanup was performed on a florisil cartridge and the purified samples were derivatized with Sylon BFT [ N,O -bis (trimethylsilyl) trifluoroacetamide (BSTFA)-trimethylchlorosilane (TMCS), 99:1, v/v] in toluene. meta -Nitrochloram- phenicol ( m -CAP) was used as an internal standard for the determination. Selected ion monitoring (SIM) was used for detection, wherein ions were chosen, respectively, for monitoring at m/z 432, 466, 468, 470 for m -CAP, m/z 376, 378, 466, 468 for CAP, m/z 409, 411, 499, 501 for TAP, and m/z 339, 341, 429, 431 for FF. Quantitative ions were selected at m/z 466 for CAP and m -CAP, at m/z 339 for FF and at m/z 409 for TAP. The detection limits were 0.03 μg/kg for CAP, 0.2 μg/kg for FF and TAP. The correlation coefficients were above 0.99 for the calibration curves of the medicines. The linear ranges were 0.1–8.0 μg/kg for CAP and 0.2–4.0 μg/kg for FF and TAP. The reproducibilities of the compounds within a batch were 5.5%, 10.4%, and 8.8% for CAP, FF and TAP respectively, while the corresponding values between the batches were 7.4%, 20.7%, and 19.1%. The recoveries were 80.0%–111.5%, and the relative standard deviations were 1.2%–15.4%. The method is applicable for detection of the residues in animal derived food, such as pork, poultry and aquatic products.

An analytical method was developed for the simultaneous determination of 12 amide herbicide residues in soybeans. The 12 amide herbicides were extracted with acetone from soybean samples. The extract was cleaned up using a Florisil cartridge, and then determined by high performance liquid chromatography. Kromasil 100-5 C18 column (4.6 mm i. d. x 250 mm, 5 microm) as analytical column, pure water-acetonitrile as mobile phase with a flow rate of 1.0 mL/min and gradient elution, and a diode array detector set at 210 nm were used. The recoveries of amide herbicides from soybeans spiked at levels of 0.05, 0.1 and 1.0 mg/kg were in the range of 75% - 102% with relative standard deviations of 1.9% - 16.1%. The method has good linear in the range of 0.05 - 1.0 mg/L for each analyte (nu0.999). The detection limit for each amide herbicide was less than 0.05 mg/kg. The method is simple, rapid and accurate. It can be used for the inspection of amide herbicide residues in soybeans.

Background Cystic echinococcosis (CE), caused by Echinococcus granulosus metacestode, invokes a serious public health concern. Early diagnosis has great impacts on reduction of disability-adjusted life years. Several antigen B-related molecules (EgAgB; EgAgB1-5) are known to be immunopotent, but detection of EgAgB is variable in many patients and may not allow reliable interpretation of its immunological relevance. More importantly, the immunoproteome profile of hydatid fluid (HF) has not been addressed. Methods We conducted a proteome analysis of the HF of a single fertile cyst of CE1 and CE2 stages through two-dimensional electrophoresis (2-DE). Each protein spot was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). We subsequently determined the immunoproteome profile employing patient sera of entire disease spectrum from CE1 to CE5 stages. Results We identified 40 parasite proteins, of which EgAgB (28 spots) and antigen 5 (EgAg5; 5 molecules) were abundant. EgAgB proteoforms constituted the majority, mostly EgAgB1 (24 spots), followed by EgAgB2 and EgAgB4 (2 spots each). EgAgB3 was detected only by liquid chromatography-MS/MS. EgAgB5 was not recognized. We also detected 38 host proteins, which were largely composed of serum components, antioxidant/xenobiotic enzymes, and enzymes involved in carbohydrate metabolism. CE1 and CE2 HF exhibited comparable spotting patterns, but CE2 HF harbored greater amounts of EgAgB and EgAg5 complexes. CE sera demonstrated complicated immune recognition patterns according to the disease progression; CE2 and CE3 stages exhibited strong antibody responses against diverse EgAgB and EgAg5 proteoforms, while CE1, CE4, and CE5 stages mainly reacted to EgAg5 and cathepsin B. Patient sera of alveolar echinococcosis (AE) cross-reacted with diverse EgAgB isoforms (36%). EgAg5 and cathepsin B also demonstrated cross-reactions with sera from neurocysticercosis and sparganosis. Conclusions Our results demonstrated that detection of a single defined molecule may not properly diagnose CE, since specific immunodominant epitopes changed as the disease progresses. Immunoproteome analysis combined with imaging studies may be practical in the differential diagnosis of CE from AE and other cystic lesions, as well as for staging CE, which are pertinent to establish appropriate patient management. Electronic supplementary material The online version of this article (doi:10.1186/s13071-014-0610-7) contains supplementary material, which is available to authorized users.