不同剂量钼对小尾寒羊组织病理变化及抗氧化功能影响

为了研究不同剂量钼对小尾寒羊病理组织学变化及抗氧化功能的影响,试验选取18只2月龄健康、大小匀称[(20±1.34)kg]小尾寒羊公羊,随机分为3组,因加钼量的多少分为Ⅰ组[60 mg/(kgBW.d)]、Ⅱ组[30 mg/(kgBW.d)]、Ⅲ组[0 mg/(kgBW.d)],每日记录临床表现,定期抽取血液样品,末期采取组织进行研究。结果表明:①两个水平的钼添加剂量下,试验羊均出现典型的钼中毒症状,剂量越高,症状表现越严重;同时,肝脏、脾脏、睾丸、心肌、肾脏均出现不同程度的病理学变化,剂量越高,病理学变化越明显,而肺脏、淋巴结无明显变化。②血清GSH-Px、SOD活性随钼剂量的增加而降低(P<0.05),血清XOD活力和血清MDA浓度随钼剂量的增加而升高(P<0.05),有明显的剂量-效应关系。研究表明,30 mg/(kgBW.d)和60 mg/(kgBW.d)钼能引起小尾寒羊组织损伤和抗氧化机能降低,钼剂量越高,毒性作用越明显。     

A multipronged approach for the detection of leptospirosis in captive sloth bears (Melursus ursinus) in Agra and Bannerghatta sloth bear rescue centers in India

Leptospirosis is an exacerbating factor responsible for the drastic decline of sloth bear population in India. In this study, a multipronged approach based on antigen detection using Polymerase Chain Reaction (PCR) employing G1/G2 and LigBF/LigBR primers, antibody detection using Microscopic Agglutination Test (MAT) and recombinant LigBCon1-5 antigen based Latex Agglutination Test (rLigBCon1-5 LAT), serum biochemistry using hepatic (serum glutamate oxalo acetic transaminase (SGOT) and serum glutamate pyruvic transaminase (SGPT) and renal biomarkers (blood urea nitrogen (BUN) and Creatinine) and gross/histopathological evidence in liver and kidneys were employed to investigate leptospirosis in captive sloth bears. A total of 133 serum samples collected from Agra (n=113) and Bannerghatta (n=20) sloth bear rescue centers were screened using MAT and rLigBCon1-5 LAT. A total of 87 and 78 sera tested positive by MAT and LAT respectively. Pyrogenes was the leading serovar obtained using MAT followed by Icterohaemorrhagiae, Javanica, Grippotyphosa, Canicola and Tarassovi. The relative sensitivity, specificity and accuracy of rLigBCon1-5 LAT in comparison to MAT were 89.66%, 100% and 93.23% respectively. PCR performed on hepatic and renal tissues showed amplicon of 285 and 219 base pairs for G1/G2 and LigBF/LigBR primers respectively. Gross evidence (icteric liver, severely engorged hepatic sinusoids, congested kidneys with necrotic white spots on sub capsular surface), histopathology (severe hepatic degeneration and tubulointerstitial nephritis) and elevated hepatic/renal biomarkers were suggestive of leptospirosis. This study suggests that rLigBCon1-5 LAT can be employed as a pen-side test for detecting leptospirosis in sloth bears.     

Basal cell tumor or cutaneous basilar epithelial neoplasm? Rethinking the cytologic diagnosis of basal cell tumors

A 1-cm-diameter, red, raised, cutaneous mass over the dorsal surface of the left third metacarpal of a 6-year-old neutered male yellow Labrador Retriever was aspirated. The aspirate contained cohesive clusters of cells, some containing cells with increased pleomorphism. Cellular debris (some keratinized), melanin, large numbers of macrophages, a few neutrophils, and fibroblasts were also observed. The cytologic interpretation was malignant neoplasia with histiocytic inflammation. Differentials included a carcinoma or, given the melanin pigment and variable morphology of the cells, possibly malignant melanoma. Histologically, the tumor was diagnosed as a basal cell epithelioma. Neoplasms that once were lumped into the broad histologic diagnosis of basal cell tumors have since been split into distinct entities, dependent on evidence of differentiation into epidermis, trichofollicular epithelium, or sweat or sebaceous glands. Although histologic reclassification has resulted in removal of most of these entities from the original basal cell tumor category, a cytologic diagnosis of basal cell tumor continues to be used to represent the large, heterogeneous group of epidermal, trichofollicular, and adnexal skin tumors with basal cell characteristics. The case in this report demonstrates the heterogeneity of neoplasms that may be diagnosed cytologically as basal cell tumors and supports the need for cytologic criteria and nomenclature that better reflect potential variation in tissue differentiation.     

Canine eyelid granular cell tumor: a report of eight cases

To describe a previously unreported neoplasm of the medial canthus and eyelid in dogs. Clinical and pathologic features of granular cell tumors in the dog were reviewed. Granular cell tumors, arising from the medial canthal eyelid of eight dogs, were identified from the archives of the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW). The affected dogs ranged in age from 5 to 12 years (mean of 9.25 years). Follow‐up information was available for seven of the eight cases. The clinical presentation included swollen hyperemic lids (4/8), ulcerated skin overlying the mass (2/8), and red conjunctiva (7/8). All eight of the cases had firm masses extending from the palpebral conjunctiva to the eyelid margin at the medial canthus. Histologically, the tissue was composed of a highly collagenous neoplastic growth. The neoplastic cells were oval to strap‐like cells with an oval bland appearing nucleus and abundant amounts of granular cytoplasm with very distinct cell boundaries. These granular cells were embedded in a dense collagen matrix. A PAS stain faintly highlighted the granular appearance of the neoplastic cells, which is a defining characteristic of this tumor. There was no recurrence in the seven cases available for follow‐up. Canine granular cell tumors of the medial canthus present clinically and histologically as a benign neoplasm. Granular cell tumors have a characteristic histological appearance. Granular cell tumors should be on the differential list for nodules of the medial canthus in dogs.     

鸭黄病毒感染病例的病理组织学观察

为了探明湖北省肉种鸭和蛋鸭产蛋下降的原因,对引起产蛋下降的病鸭的卵巢中分离的病毒的目的基因进行测序,对3代接种的SPF鸡胚尿囊液进行电镜观察,对脑等组织进行组织病理学观察。序列分析结果表明,扩增的序列属于黄病毒的NS5基因,且与恩塔亚病毒群有相近的遗传进化关系。电镜可观察到40nm～60nm的有囊膜的病毒粒子。组织病理学观察发现输卵管出现严重的充血、出血和炎性反应,脑充血、炎性反应,心脏中有大量变性坏死的炎性细胞。     

Equine laminitis model: Lamellar histopathology seven days after induction with oligofructose

Reasons for performing study: The histopathology of laminitis during its transition from the acute to the chronic phase has not been previously documented. Studying hoof lamellar tissues 7 days after induction of laminitis may provide insight into the intractable nature of the chronic phase of the disease. Objectives: To induce laminitis and investigate hoof wall lamellar tissues 7 days after dosing. Methods: Laminitis was induced using oligofructose in 6 normal Standardbred horses. The dorsal hoof lamellar tissues of these and 12 normal horses were processed and examined by light microscopy. Serial sections of a lamellar tip affected by laminitis were used to create a 3 dimensional reconstruction. Results: Transverse sections of dorsal hoof wall lamellae were significantly longer than normal. Many secondary epidermal lamellae were not connected to primary lamellae and existed as spherical or ovoid, discrete islands isolated in the lamellar dermis. The lamellar basement membrane was intact. Conclusions: Lamellar tissue has the ability to reorganise rapidly following an episode of acute laminitis. Although histopathological evidence of ongoing acute laminitis was absent by 7 days, there was marked disruption of lamellar architecture. Potential relevance: The architecture and subsequent strength of the resultant lamellar interface could be greatly influenced for the better by strategies that minimise mechanical displacement during the acute phase of laminitis.     

Segmentation of the Pathophysiological Stages of Diabetic Changes in the db/db Mouse

The db/db mouse is one of the diabetes mellitus animal models and if the pathophysiological stages of diabetic changes in the mouse model could simulate the stages in human diabetes, the db/db mouse could be used to better evaluate drug candidates. Blood insulin, HbA1c levels and morphological features of pancreatic islets in db/db mice were evaluated to determine the pathophysiological stage. At 6 weeks of age, db/db mice showed the highest level of plasma insulin and lowest level of HbA1c, and histopathological examination revealed enlarged islets with a circular shape and hypertrophic islet cells. By 9 and 12 weeks of age, the plasma insulin levels had decreased to mid levels and HbA1c had increased to mid to high levels; histopathological examination at this time revealed two types of islets coexisting, enlarged circular islets and small irregular-shaped islets. By 15 and 22 weeks of age, plasma insulin had decreased further to low levels and HbA1c was at its highest level; the histopathological examination at this time revealed an increase in irregular-shaped and small islets. Based on blood insulin levels, HbA1c levels and histopathology findings in the db/db mice in this study, the clinical staging of diabetic changes were recognized. The pathophysiological stages of diabetes mellitus in this animal model were similar to the stages in humans.     

鸭瘟病毒对SPF鸭的致病性研究

【目的】探讨鸭瘟病毒（Duck plague virus,DPV）对SPF鸭的致病性。【方法】将DPV分别通过点眼滴鼻和皮下注射接种途径人工感染SPF鸭,以未攻毒健康SPF鸭为对照,观察各组鸭的发病情况及症状,于感染后不同时间随机剖杀试验组和对照组SPF鸭,采集肝脏、脾脏、肾脏、肺、心脏、气管、食道、肠道、胸腺、法氏囊、脑和泄殖腔等组织,常规方法制备石蜡切片,HE染色,观察各器官病理组织学变化,同时利用间接免疫荧光染色法（IFA）对DPV抗原在鸭体内的分布进行检测,并进行血常规和血液生化指标检测,分析了DPV对SPF鸭淋巴细胞转化率的影响。【结果】DPV接种后,引起鸭高热、下痢,组织器官出血、坏死,食道、泄殖腔黏膜出血、溃疡并有灰黄色假膜覆盖;病理组织学变化以血管壁损伤为主,肝、脾细胞变性、坏死,消化道黏膜上皮细胞坏死脱落。DPV感染后,在鸭的脾脏、胸腺、法氏囊、肝脏、食道、泄殖腔、肠道、肾脏、肺及气管中均检测到DPV抗原,且以肝脏、脾脏等组织中荧光最强;在脑和心脏中均未检测到DPV抗原。DPV感染组鸭血清中白细胞总数（WBC）、红细胞总数（RBC）、血小板总数（PLT）和总蛋白（TP）、血红蛋白（HGB）含量及丙氨酸氨基转移酶（ALT）、碱性磷酸酶（ALP）、天门冬氨酸氨基转移酶（AST）活性、淋巴细胞转化率与对照组差异大多达显著或极显著水平。【结论】DPV感染主要引起SPF鸭各组织器官广泛性出血;脾脏、胸腺、法氏囊等免疫器官首先受到攻击和损害,并造成免疫抑制;皮下注射接种DPV比点眼滴鼻接种对SPF鸭的致病性强。     

Detection of Mycobacterium avium subsp paratuberculosis in formalin-fixed paraffin-embedded intestinal tissue by IS900 polymerase chain reaction

OBJECTIVES: To evaluate and compare methods for DNA extraction from formalin-fixed, paraffin-embedded tissues and methods for detection of Mycobacterium avium subsp paratuberculosis by IS900 PCR for confirmation of Johne's disease in ruminants. DESIGN: A laboratory study. PROCEDURE: Three methods of DNA extraction of differing complexity and two PCR protocols using different pairs of IS900 primers were compared. Sensitivity and specificity were assessed using samples from ruminants with and without histological evidence of Johne's disease. RESULTS: The simplest method of DNA extraction, which involved two cycles of boiling and freezing followed by centrifugation, gave more consistent results than two methods that required solvent extraction of paraffin, proteinase digestion and DNA purification. The sensitivity of detection of M avium subsp paratuberculosis in paraffin blocks stored for 1 to 6 years from 34 cases of Johne's disease in sheep, cattle and goats was 88% for a 229 bp IS900 PCR assay and 71% for a 413 bp assay, using the detection of acid-fast bacilli by Ziehl Neelsen staining of histological sections from the same blocks as the gold standard test. PCR results correlated with the abundance of acid-fast organisms in the tissues. No false positive reactions were detected. CONCLUSION: PCR for identification of M avium subsp paratuberculosis in formalin-fixed, paraffin-embedded intestinal tissues from ruminants is a rapid and useful method. A simple method of sample preparation is effective. Amplification of short fragments of IS900 is more effective than amplification of longer fragments.     

Sublethal Toxicity of the Herbicide Butachlor on the Earthworm Perionyx sansibaricus and its Histological Changes (5 pp)

Goal, Scope and Background   With the advent of the Green Revolution, there has been a quantum leap in the use of synthetic herbicides and pesticides throughout the world to sustain high-yielding crop varieties. Continuous use of these synthetic chemicals leads to loss of soil fertility and soil organisms. Histopathological studies may signal a damaging effect of organisms resulting from prior or ongoing exposure to toxic agents. A large number of studies have reported general histological changes in earthworms. A fewer studies have reported more specific types of histopathological studies in Eisenia foetida, Dendrodrilus rubidus, Lumbricus terrestris, Lumbricus rubellus and Octolasium transpandanum. However, no reports are available on Perionyx sansibaricus. The aim of the present study is (i) to determine the sublethal effects of the herbicide Butachlor on Perionyx sansibaricus on growth and cocoon production, (ii) to evaluate the histological changes in the exposed worms.Methods   The test substrate, namely powdered, dried cow dung, was contaminated with 0.0657, 0.1315 and 0.1973 mg kg–1 of Butachlor, and a control was maintained. Four groups of each 10-test species of Perionyx sansibaricus were used per concentration level (control, 0.0657, 0.1315 and 0.1973 mg kg–1). Various life history parameters such as biomass, cocoon production and histological changes were measured.Results   Worm growth was observed at various exposures over 60 days. While exposing the earthworm to the herbicide, no mortality was observed, not even at a higher dose. At the end of the experiment, the control group had a mean biomass of 0.0831±0.00 mg and, in the exposed group, at herbicide concentrations of 0.1973, 0.1315 and 0.0657 mg kg–1, the mean biomass was found to be 0.0497±0.00 mg, 0.0628±0.00 mg and 0.0781±0.00 mg, respectively. The mean earthworm biomass was found to be decreased with increasing herbicide concentration. Similarly, cocoon production was also reduced by the increasing herbicide concentration. All earthworms in the exposed group were found to have glandular cell enlargement and to be vacuolated. Conclusion   The results clearly indicate that Butachlor can retard the growth and cocoon production in Perionyx sansibaricus. The effect of the herbicide Butachlor can be assessed by the histological observation of the intestinal region, as evidenced by glandular cell enlargement at all the exposed concentrations, which may massively affect food intake and which in turn may indirectly inhibit the earthworm reproductive capacity.Recommendation and Outlook   Risk assessment is normally aimed at the protection of human health and the ecosystem, and the interrelationship of these two areas of protection is easy to perceive. The use of earthworms in risk assessment is to obtain more information on environmental quality and ensure environmental safety. There is an urgent need to test the chemicals causing toxicity to earthworms, because earthworms play a major role in soil fertility as well as acting as a transferring route from the soil to the terrestrial ecosystem.