New multiplex PCR method for the simultaneous diagnosis of the three known species of equine tapeworm

Although several techniques exist for the detection of equine tapeworms in serum and feces, the differential diagnosis of tapeworm infection is usually based on postmortem findings and the morphological identification of eggs in feces. In this study, a multiplex polymerase chain reaction (PCR)-based method for the simultaneuos detection of Anoplocephala magna, Anoplocephala perfoliata and Anoplocephaloides mamillana has been developed and validated. The method simultaneously amplifies hypervariable SSUrRNA gene regions in the three tapeworm species in a single reaction using three pairs of primers, which exclusively amplify the internal transcribed spacer 2 (ITS-2) in each target gene. The method was tested on three types of sample: (a) 1/10, 1/100, 1/500, 1/1000, 1/2000 and 1/5000 dilutions of 70 ng of genomic DNA of the three tapeworm species, (b) DNA extracted from negative aliquots of sediments of negative control fecal samples spiked with 500, 200, 100, 50 and 10 eggs (only for A. magna and A. perfoliata; no A. mamillana eggs available) and (c) DNA extracted from 80, 50, 40, 30, 10 and 1 egg per 2 μl of PCR reaction mix (only for A. magna and A. perfoliata; no A. mamillana eggs available). No amplification was observed against the DNA of Gasterophilus intestinalis, Parascaris equorum and Strongylus vulgaris. The multiplex PCR method emerged as specific for the three tapeworms and was able to identify as few as 50 eggs per fecal sample and as little as 0.7 ng of control genomic DNA obtained from the three species. The method proposed is able to differentiate infections caused by the two most frequent species A. magna or A. perfoliata when the eggs are present in feces and is also able to detect mixed infections by the three cestode species.     

一种蓝莓不良症状的诊断方法

本文介绍了一种分析蓝莓生长不正常时原因的方法,通过4个步骤的原因分析,从而帮助蓝莓种植者作出正确地判断并解决问题。     

兔粘液瘤病诊断方法的研究

注射过粘液瘤病毒的兔子全部死亡,主要病变为皮下发生粘液性肿胀,和真皮中有星形、梭形或蝌蚪形瘤细胞浸润,在表皮的基细胞和棘细胞层中及瘤细胞中存在有不同阶段的病毒粒子和病毒包涵体,建立了琼脂扩散检测法用于粘液瘤病的诊断。     

我国兽医诊断制品管理概况与监管建议

为提高兽医诊断制品的有效供应和质量，全面总结国内兽医诊断制品现状，对比研究国内外诊断制品管理情况，分析当前国内兽医诊断制品存在的主要问题，并有针对性地提出了监管建议。     

Plasma Procalcitonin Concentration in Healthy Horses and Horses Affected by Systemic Inflammatory Response Syndrome

Background

The diseases most frequent associated with SIRS in adult horses are those involving the gastrointestinal tract. An early diagnosis should be the goal in the management of horses with SIRS.

Objective

The objective of this study was to evaluate the plasma procalcitonin (PCT) concentration in healthy and SIRS horses to assess differences between the two groups.

Animals

Seventy‐eight horses (30 healthy and 48 SIRS).

Methods

Prospective in vivo multicentric study. Horses were classified as SIRS if at least 2 of the following criteria were met: abnormal leukocyte count or distribution, hyperthermia or hypothermia, tachycardia, tachypnea. Healthy horses showed no clinical or laboratory signs of SIRS. Plasma PCT concentrations were measured with a commercial ELISA assay for equine species. Results were expressed as mean±standard deviation. T‐test for unpaired data was performed between healthy and SIRS group. SIRS group was divided in 4 subgroups and t‐test was performed between healthy versus each subgroup.

Results

PCT concentrations in healthy and SIRS horses were 18.28 ± 20.32 and 197.0 ± 117.0 pg/mL, respectively. T‐test showed statistical differences between healthy versus SIRS group and between healthy versus all subgroups.

Conclusions and Clinical Importance

Results showed an increase in PCT concentration in SIRS horses as previously reported in humans and dogs. PCT could be used as a single assay in equine practice for detection of SIRS.     

鸡传染性鼻炎实验室诊断技术

对鸡传染性鼻炎病研究作了简要介绍，并结合本实验室的实际工作经验，重点介绍了本病实验室诊断技术。     

Mitral valve morphology assessed by three-dimensional transthoracic echocardiography in healthy dogs and dogs with myxomatous mitral valve disease

Objective

To assess differences in morphology of the mitral valve (MV) between healthy dogs and dogs affected by myxomatous mitral valve disease (MMVD) using real-time transthoracic three-dimensional echocardiography (RT3DE).

区分禽流感病毒亚型诊断基因芯片的构建

制备了可同时区分AIVH5、H7、H9血凝素亚型及N1、N2神经氨酸酶亚型的基因诊断芯片。试验以重组质粒为模板,进行PCR扩增,然后用异丙醇沉淀法进行纯化,制备探针及内参基因。将探针及内参基因用点样缓冲液稀释到0.3μg/μL后用芯片点样仪将其点制到醛基化基片上,样点中心间距450μm,样点直径220μm。将点好的基片在室温干燥24h以上,后经65℃再水合10s、80℃干燥、65mj紫外线交联照射25min后,再用封闭液封闭,95℃变性处理,成功构建了能区分禽流感血凝素H5,H7,H9亚型及神经氨酸酶N1,N2亚型的检测基因芯片。在PCR扩增中标记待检样品,对制备的检测芯片进行质量检验,结果表明,制备的区分禽流感病毒亚型基因芯片可区分AIV部分亚型。     

Agreement of Serum Spec cPL with the 1,2‐o‐Dilauryl‐Rac‐Glycero Glutaric Acid‐(6′‐methylresorufin) Ester (DGGR) Lipase Assay and with Pancreatic Ultrasonography in Dogs with Suspected Pancreatitis

Background

Spec cPL is the most sensitive and specific test for diagnosing pancreatitis in dogs. Its results have not been compared to those of the 1,2‐o‐dilauryl‐rac‐glycero‐3‐glutaric acid‐(6′‐methylresorufin) ester (DGGR) lipase assay or those of abdominal ultrasonography.

Objectives

To investigate agreement of Spec cPL with DGGR lipase activity and pancreatic ultrasonography in dogs with suspected pancreatitis.

Animals

One hundred and forty‐two dogs.

Methods

DGGR lipase activity (reference range, 24–108 U/L) and Spec cPL were measured using the same sample. The time interval between ultrasonography and lipase determinations was <24 hours. The agreement of the 2 lipase assays at different cutoffs and the agreement between pancreatic ultrasonography and the 2 tests were assessed using Cohen''s kappa coefficient (κ).

Results

DGGR lipase (>108, >216 U/L) and Spec cPL (>200 μg/L) had κ values of 0.79 (95% confidence interval [CI], 0.69–0.9) and 0.70 (CI, 0.58–0.82). DGGR lipase (>108, >216 U/L) and Spec cPL (>400 μg/L) had κ values of 0.55 (CI, 0.43–0.67) and κ of 0.80 (CI, 0.71–0.9). An ultrasonographic diagnosis of pancreatitis and DGGR lipase (>108, >216 U/L) had κ values of 0.29 (CI, 0.14–0.44) and 0.35 (CI, 0.18–0.52). Ultrasonographically diagnosed pancreatitis and Spec cPL (>200, >400 μg/L) had κ values of 0.25 (CI, 0.08–0.41) and 0.27 (CI, 0.09–0.45).

Conclusions and Clinical Importance

Although both lipase assays showed high agreement, agreement between ultrasonography and lipase assays results was only fair. Because lipase results are deemed more accurate, ultrasonography results should be interpreted carefully.     

奶牛乳腺炎诊断方法研究进展

奶牛乳腺炎的诊断方法有很多,临床乳腺炎可根据临床症状进行诊断,用于隐性乳腺炎的诊断方法有乳汁细胞学检查、乳汁的细菌培养、电导率测定、pH值的检查、乳清电泳诊断法、酶检测法、PCR检测方法、被毛微量元素检验法以及其它衍生方法.本文对奶牛乳房炎诊断研究进展作了系统的介绍.     

Evaluation of static telepathology in veterinary diagnostic cytology

BACKGROUND: Telepathology is the process of diagnostic pathology performed on digital images viewed on a display screen rather than by conventional light microscopy with glass slides. At present, electronic mail (e-mail) attachment is the most common communication medium for telepathology, because it is relatively simple and incurs minimal cost. OBJECTIVE: Our objective was to test whether telediagnosis through Internet e-mail could be applied to veterinary diagnostic cytology. METHODS: Using 20 cytology cases, on which a consensus diagnosis was reached by 3 experienced pathologists, a total of 130 images were digitized and sent as e-mail attachments from the Unit of Anatomic Pathology of the Faculty of Veterinary Medicine of Naples to the Unit of Pathology of the Faculty of Veterinary Medicine of Messina, Italy. The images were reviewed at the Unit of Pathology of Messina, where the consulting pathologist formulated his diagnoses by visualizing the images on the monitor of a computer. The telediagnoses were compared with the consensus diagnoses and with the consulting pathologist's interpretation using conventional light microscopy at a later date. RESULTS: The median time to capture images was approximately 30 minutes for each case. The median time to make a telecytodiagnosis was a few seconds for each case. Overall, there was good agreement (85%) between the consensus diagnosis and the consultant's telediagnosis. In 100% of the cases there was agreement between the consulting pathologist's telediagnosis and conventional glass slide diagnosis. CONCLUSION: Telepathology by e-mail provides acceptable efficacy and a faster turnaround time than post and can be applied to veterinary diagnostic cytology.     

鸡传染性法氏囊病（IBD）琼脂扩散试验（AGP）诊断试剂的研制

用典型发病鸡的法氏囊制备抗原,以免疫鸡卵黄抗体代替阳性对照血清,研制出鸡传染性法氏囊病（ＩＢＤ）琼脂扩散试验（ＡＧＰ）诊断试剂,用于检测免疫鸡群的ＩＢＤ抗体,指导疫苗免疫;检测发病鸡群的法氏囊组织ＩＢＤ抗原,达到确诊     

Infrared Thermography in Dogs with Mammary Tumors and Healthy Dogs

Background

Infrared thermography is a painless, noninvasive, nonionizing diagnostic imaging exam used in human medicine as an auxiliary tool for breast cancer diagnosis in women.

Hypothesis/Objectives

Define thermographic mean temperatures of healthy mammary glands and compare these temperatures with those of mammary glands with tumors in dogs.

Animals

Fifty client‐owned female dogs were evaluated, including 20 with histopathologically confirmed mammary tumor and 30 clinically healthy (control).

Methods

A randomized study using infrared thermography analyzed each mammary gland of the animals from the control group and mammary glands with tumors from the tumor group, then the thermographic temperatures obtained were compared. Thermographic exam was performed in a temperature‐controlled room with a cooled thermographic camera—Flir E‐40 (Flir Systems^®)

Results

There was significantly a higher temperature in the caudal abdominal and inguinal mammary glands than the other glands in the healthy group (P < .05). Dogs with mammary tumors had significantly higher thermographic temperature compared with unaffected glands regardless of the tumor size and the location (P < .05).

Conclusions and clinical importance

The technique seems to be able to assess for the presence of neoplasia within the mammary tissue in bitches. Further investigation is necessary to determine the impact of this technique when adopted clinically.