Assessment of Aspergillus fumigatus burden in lungs of intratracheally-challenged turkeys (Meleagris gallopavo) by quantitative PCR,galactomannan enzyme immunoassay,and quantitative culture

Aspergillus fumigatus remains a major respiratory pathogen in birds and treatment is still difficult. We challenged different groups of few-day-old turkeys via intratracheal aerosolisation with increasing concentrations (10⁵ up to 10⁸) of conidia using a MicroSprayer^® device. The fungal burden was assessed by real-time PCR, galactomannan dosage, CFU counting and histopathological evaluation in order to provide a comparison of these results within each inoculum groups. Significant mortality, occurring in the first 96 h after inoculation, was only observed at the highest inoculum dose. Culture counts, GM index and qPCR results on the one hand and inoculum size on the other hand appeared to be clearly correlated. The mean fungal burden detected by qPCR was 1.3 log₁₀ units higher than the mean values obtained by CFU measurement. The new model and the markers will be used to evaluate the efficacy of antifungal treatments that could be used in poultry farms.     

Agar/galactomannan blends for strawberry (Fragaria x ananassa Duchesne) cv. Pelican micropropagation

The efficacy of partial substitution of agar by galactomannans (GMs) obtained from different sources was tested in strawberry (Fragaria x ananassa Duchesne) cv. Pelican micropropagation. GMs extracted from seeds of Cassia fastuosa (cassia) and Cyamopsis tetragonolobus (guar gum – a commercial GM) were mixed with agar in the proportion of 0.3/0.3% (w/v) in MS medium, and the performance was compared with behaviour of medium containing agar only (0.6%, w/v) as gelling agent. The results obtained showed that agar could be partially replaced by GMs, since the rheological measurements confirmed a good interaction between the polysaccharides. Also, better performance and enhanced proliferation of strawberry shoots were observed; where in the modified medium the length of roots was higher. Thus, the behaviour of the agar/galactomannans gels and the commercial cost effectiveness, which is lower for the modified medium compared with the system solidified with agar only, showed the potential use of these galactomannans for application in plant tissue culture.     

槐树的化学成分及其利用

本文介绍了槐树的形态特征、生长环境、化学成分和用途。     

Clinical Features and Magnetic Resonance Imaging Findings in 7 Dogs with Central Nervous System Aspergillosis

Background

Systemic aspergillosis is a manifestation of Aspergillus sp. infection that can result in central nervous system (CNS) involvement with marked alterations in CNS function. Information regarding the clinical presentation and magnetic resonance imaging (MRI) findings in cases of aspergillosis with CNS involvement is lacking, resulting in a need for better understanding of this disease.

Hypothesis/Objectives

The primary objectives were to describe the clinical features and MRI findings in dogs with CNS aspergillosis. The secondary objectives were to describe clinicopathologic findings and case outcome.

Animals

Seven dogs with CNS aspergillosis.

Methods

Archived records from 6 institutions were reviewed to identify cases with MRI of CNS aspergillosis confirmed with serum galactomannan enzyme immunoassay (EIA) testing, culture, or supported by histopathology. Signalment, clinical, MRI, clinicopathologic, histopathologic, and microbiologic findings were recorded and evaluated.

Results

Aspergillosis of the CNS was identified in 7 dogs from 3 institutions. The median age was 3 years and six were German Shepherd dogs. Five dogs had signs of vestibular dysfunction as a component of multifocal neurological abnormalities. The MRI findings ranged from normal to abnormal, including hemorrhagic infarction and mass lesions.

Conclusions and Clinical Importance

Until now, all reported MRI findings in dogs with CNS aspergillosis have been abnormal. We document that CNS aspergillosis in dogs, particularly German Shepherd dogs, can be suspected based on neurologic signs, whether MRI findings are normal or abnormal. Confirmatory testing with galactomannan EIA, urine, cerebrospinal fluid (CSF) or tissue culture should be performed in cases where aspergillosis is a differential diagnosis.     

微水固相法氧化改性皂荚多糖胶

采用微水固相法氧化改性皂荚多糖胶(GSG),以H2O2为氧化剂,制备氧化皂荚多糖胶(OGSG)。通过凝胶渗透谱(GPC)和黏度计检测器等的分析,比较不同质量分数的H2O2对OGSG的取代度(DS)、相对分子质量、黏度的影响。结果显示:随着氧化试剂H2O2浓度的增大,氧化皂荚多糖胶的取代度和多分散性呈增加趋势,特性黏度和相对分子质量呈下降趋势。皂荚多糖胶20 g,30%H2O218 g,反应时间1 h,反应温度65℃,在上述较佳反应条件下,取代度达到1.22。微水固相法氧化改性可以在使用较少反应试剂条件下达到较好氧化的效果。     

Evaluation of serum galactomannan detection for diagnosis of feline upper respiratory tract aspergillosis

Measurement of serum galactomannan (GM), a polysaccharide fungal cell-wall component, is a non-invasive test for early diagnosis of invasive aspergillosis in humans. Feline upper respiratory tract (URT) aspergillosis is an emerging infectious disease in cats. Diagnosis requires biopsy for procurement of tissue specimens for cytological or histological detection of fungal hyphae and for fungal culture. The aim of this study was to evaluate serum GM measurement as a non-invasive diagnostic test for URT aspergillosis in cats. A one-stage, immunoenzymatic sandwich ELISA was used to detect serum GM in 4 groups of cats; Group 1 (URT aspergillosis) – confirmed URT aspergillosis (n = 13, sinonasal aspergillosis (SNA) n = 6 and sino-orbital aspergillosis (SOA) n = 7), Group 2 (URT other) – other URT diseases (n = 15), Group 3 (β-lactam) – cats treated with β-lactam antibiotics for non-respiratory tract disease (n = 14), Group 4a – healthy young cats (≤1 y of age, n = 28), Group 4b – healthy adult cats (>1 y of age, n = 16). One cat with SNA and two cats with SOA caused by an Aspergillus fumigatus-mimetic species, tested positive for serum GM. For a cut-off optical density index of 1.5, the overall sensitivity and specificity of the assay was 23% and 78% respectively. False positive results occurred in 29% of cats in Group 3 and 32% of cats in Group 4a. Specificity increased to 90% when Groups 3 and 4a were excluded from the analysis. Overall, serum GM measurement has a poor sensitivity but is a moderately specific, non-invasive screening test to rule out infection in patients with suspected feline upper respiratory tract aspergillosis.     

瓜尔豆发育过程中碳水化合物含量的变化

研究了瓜尔豆生长发育过程中叶片、种荚及籽粒中蔗糖合成酶（SS）和蔗糖磷酸合成酶（SPS）活性的变化、糖类物质的组成及其含量的动态变化,以揭示可溶性糖的合成对瓜尔豆胚乳中半乳甘露聚糖和淀粉积累的影响。结果表明,进入开花期（播种后45 d）后,叶片、种荚和籽粒中SS和SPS活性均呈降低趋势。籽粒中的SS和SPS活性呈正相关,且显著高于叶片和种荚中相应酶活性。叶片中葡萄糖（Glc）含量高于蔗糖（Suc）。种荚中可溶性糖含量与Suc含量均呈单峰曲线,在籽粒直径5 mm时达峰值,两者呈正相关。Suc是种荚中暂存含量最多的可溶性糖,但其含量与SS和SPS活性无显著相关性,它主要来自叶片。种荚中Suc含量与籽粒碳水化合物总量（可溶性糖+多糖）呈负相关,它是籽粒碳水化合物合成的重要物质来源,还与籽粒库的建成有关。随种子发育,籽粒可溶性糖含量呈逐步下降趋势。籽粒发育前期,叶片同化物以Suc的形式运输并大量积累在种荚中,当籽粒直径达5 mm后迅速由种荚卸载至籽粒。但卸载期籽粒Suc含量较低,且显著低于种荚,说明籽粒Suc迅速转化为其他可溶性糖,并进一步用于多糖合成。籽粒直径为4 mm至5 mm时是种荚由“源”到“库”转换的临界期,这时籽粒中淀粉积累较迅速,半乳甘露聚糖在籽粒直径达5 mm后进入迅速积累期。     

Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Plateliat test is unreliable for the diagnosis of canine SNA.