Ochratoxin A and citrinin induced nephrosis in Beagle dogs. III. Terminal renal ultrastructural alterations

The extent and type of renal ultrastructural changes in Beagle dogs varied with the administration of ochratoxin A and citrinin alone and in the two dosage combinations. The three predominant changes were cytoplasmic vacuolation, myelin figure formation and lesions designated as cytoplasmic disarray. These changes were mainly of the endomembane system of the tubular epithelial cells. Cytoplasmic vacuoles were within proximal and distal tubules and collecting ducts and were most numerous in dogs given 10 mg/kg critrinin. Vacuolation of similar distribution, but less severe, was seen in renal tubular cells of dogs given the higher dose of the combined mycotoxins (0.2 mg/kg ochratoxin A + 10 mg/kg citrinin). This damage was limited to the proximal tubular cells in dogs given only ochratoxin A (0.1 or 0.2 mg/kg). Myelin figures were in proximal epithelial cells of dogs given ochratoxin A alone or combined with citrinin. There was cytoplasmic disarray in dogs of all groups except for dogs given 5 mg/kg citrinin. This lesions was usually limited to the proximal tubules. The lesions, however, was found in cells of the distal tubules of dogs given 10 mg/kg citrinin alone.     

Ultrastructure of the testis of Pekin ducks fed methyl mercury chloride: seminiferous epithelium

Testicular cells of Pekin ducks (Anas platyrhynchos) fed with 0 (control), 0.5 (group 1), 5 (group 2), or 15 (group 3) mg of methyl mercury chloride (CH3HgCl)/kg of basal feed for 12 weeks were examined by electron microscope. Sertoli's cells from ducks in group 2 had dilated smooth endoplasm reticulum, increased lysosomes, and large vacuoles, some with lipid droplets. Degenerative changes were more advanced in group 3 ducks. There were increases in lysosomes, myelinoid figures, vacuolations, cytoplasmic and nuclear debris, cristolyses of mitochondria, and distended Golgi's complexes, and a reduction in smooth endoplasm reticulum and microtubules when compared with those of the controls. Spermatogonia were resistant to CH3HgCl exposure, except in 2 ducks from group 3 which had cells that showed electron-lucent cytoplasm, abnormal mitochondria, and membrane-bound vacuoles. In primary spermatocytes, degenerative changes were evident in ducks fed the larger dose levels. In nuclei, synaptonemal complexes showed unpaired elements. In cytoplasm, cellular debris and vacuoles predominated. There was an increase in synchronized meiosis and apparent incomplete cell division. In ducks from group 3, the cellular damage was more severe and was present throughout the germinal epithelium. Spermatids differentiation was affected variably in groups 2 and 3. Severity of damage increased with the increased dosage of mercury. Where there was spermiogenic activity, the electron-dense acrosome granules, manchette, and midpiece were rarely found. Since the seminiferous tubules from 2 ducks in group 3 had severe destruction of spermatocytes and spermatids, the spermiogenic activity was negligible. Ingestion of CH3HgCl caused toxic injury to seminiferous tubules in groups 2 and 3 ducks. The degree of damage was related to the dietary amount of mercury.     

Spontaneous Accumulation of Globotriaosylceramide (Gb3) in Proximal Renal Tubules in an ICR Mouse

This report describes spontaneous cytoplasmic vacuolation in the proximal renal tubules of a 7-week-old male ICR [Crlj:CD1(ICR)] mouse. The contents of vacuoles were positively stained with periodic acid-Schiff (PAS) and Sudan black, and the membranes were positive on immunohistochemical staining for lysosomal-associated membrane protein-2 (LAMP-2), a marker of lysosomal membrane. Electron microscopy revealed electron-dense lamellar bodies in the proximal tubular epithelial cells. These histopathological features are similar to those in α-galactosidase A-deficient mice, in which globotriaosylceramide (Gb3), a glycosphingolipid, accumulates in lysosomes. When we performed immunohistochemical staining for Gb3, the contents of vacuoles were positively stained. From these results, spontaneous cytoplasmic vacuolation in the proximal renal tubules in the mouse was identified as lysosomal accumulation of Gb3.     

Ultrastructural alterations in furazolidone-induced cystic testicular degeneration in ducklings

Cystic testicular degeneration was induced in groups of ducklings by feeding furazolidone at 250, 400, 550, or 750 mg per kilogram of feed (ppm) for 4 weeks. In normal ducklings, tall, columnar Sertoli cells predominated in the seminiferous epithelium. In treated ducklings, the mildest ultrastructural alteration was cytoplasmic vacuolation of Sertoli cells. In birds with more severely affected tubules, cytoplasmic vacuolation was more severe and the cells were cuboidal or rounded. The rounded cells desquamated into the tubular lumens, eventually undergoing cytolysis. In birds with the most severely affected tubules, only extensively flattened epithelial cells lined the extremely dilated seminiferous tubules. The interstitium was edematous and compacted between the expanded tubules. Many of the testicular ultrastructural alterations in furazolidone-intoxicated ducklings were similar to those described in the testicles of sodium-intoxicated cockerels.     

Karnovsky’s fixative prevents artifacts appearing as vacuolation derived from tissue processing in kidneys treated with antisense oligonucleotide

Antisense oligonucleotide (ASO) therapies have been identified as a new treatment modality for intractable diseases. In kidneys treated with ASOs, vacuoles, in addition to basophilic granules, are often observed in the proximal tubules. Some reports have described that these vacuoles are likely to be a secondary phenomenon resulting from the extraction of ASOs during tissue processing. In this study, we compared renal morphology after fixation with Karnovsky’s fixative or 4% paraformaldehyde phosphate buffer (PFA) with that of 10% neutral-buffered formaldehyde solution (NBF). Female Sprague-Dawley rats, intravenously treated four times with 50 mg/kg locked nucleic acid containing antisense oligonucleotides (LNA-ASOs) for 1 or 2 weeks, were examined. Microscopically, vacuoles and basophilic granules in the proximal tubules were observed in the kidneys fixed with NBF. Basophilic granules are indicative of the accumulation of ASOs. Moreover, some of the vacuoles also contained faint basophilic granules, suggesting that the vacuoles were relevant to the accumulation of ASOs. Although moderate vacuolation was observed in the proximal tubules, the majority of the vacuolated epithelia were negative for kidney injury molecule-1 on immunohistochemical staining. Vacuoles in the proximal tubules were not observed in samples subjected to Karnovsky’s fixation, although basophilic granules were observed. In samples subjected to PFA fixation, vacuoles and basophilic granules were observed in the proximal tubules, similar to those in samples subjected to NBF fixation. Overall, our findings demonstrated the possibility of overestimation of vacuolation due to artifacts during tissue processing when using conventional NBF fixation. Karnovsky’s fixative is considered a useful alternative for distinguishing artificial vacuoles from true nephrotoxicity.     

Renal pathology in chicks following water deprivation

Total water deprivation of 2-day-old broiler chicks for 24, 48, 72, 96, and 120 hours did not result in visceral urate deposits. Renal tubular changes consisted of increased spaces between membrane infoldings of the distal convoluted tubular epithelium, increased number of cytoplasmic vacuoles in proximal convoluted tubules, and increased mucin production and dilation of collecting ducts.     

Pathogenesis of duck plague in the bursa of Fabricius, thymus, and spleen.

White Pekin ducks were inoculated orally with duck plague virus and killed at 24-hour intervals after inoculation. Spleen, thymus, and bursa of Fabricius were collected and examined by light, fluorescent, and electron microscopy. Necrosis of lymphocytes occurred in the bursa of Fabricius, thymus, splenic periarteriolar lymphoid sheath (T lymphocytes), and splenic germinal centers (B lymphocytes). Viral nucleocapsids were present in the karyoplasm of lymphocytes, but these cells necrotized before virions were formed. Periarteriolar reticular sheath cells and sinusoidal lining cells in the spleen, epithelial cells in Hassall's corpuscle of the thymus, epithelial cells between the cortex and medulla of the follicles in the bursa of Fabricius, and macrophages in all 3 tissues contained nucleocapsids in the nuclei and virions in cytoplasmic vacuoles before necrosis occurred.     

Nephron segment identification in the normal canine kidney by using lectin histochemistry

Lectin-binding patterns in normal canine kidneys were histochemically investigated using eight lectins. WGA, ConA, and RCA-I showed positive signals in glomerular capillary walls, with signals for RCA-I being detected heterogeneously. In tubular segments, signals for WGA, s-WGA, ConA, and RCA-I were distributed widely from proximal tubules to collecting ducts, whereas those for SBA, PNA, DBA, and UEA-I were localized in thin limbs of the loop of Henle, thick ascending limbs, distal tubules, or collecting ducts. Apart from PNA and UEA-I, lectins showed heterogeneous bindings in collecting ducts with the heterogeneity. UEA-I-positive reactions were restricted to those parts of the distal tubules in close proximity to the glomeruli, and in these parts, signals in the macula densa were markedly stronger than in other regions. Based on the present findings, lectin probes, singly or in combination, could be utilized to identify the affected nephron segment in canine renal pathology.     

Renal Tubular Cyst Formation in Newborn Rats Treated with p-Cumylphenol

In this study, we investigated the sequential changes in the development of renal tubular cysts in newborn rats treated with p-cumylphenol (PCP). Fifteen animals per sex were treated orally with 300 mg/kg/day of PCP for up to 18 days from postnatal day (PND) 4 and were sacrificed on PNDs 8, 12, 19 and 22 and after a 7 day recovery period. On PNDs 8 and 12, slight dilatation of the collecting ducts was frequently observed in the medulla and slight papillary necrosis was also noted in some cases. These dilated collecting ducts were lined with slightly hyperplastic epithelial cells. On PNDs 19 and 22, multiple large cystic changes arising from the collecting ducts in the outer medulla were seen. These cystically dilated ducts were also lined with hyperplastic epithelial cells. During the dosing period, the labeling index of proliferating cell nuclear antigen in the collecting duct epithelium was higher in the PCP-treated group than in the control group at all time points. After a 7 day recovery period, the cystic change still remained, although the cell density was decreased and the epithelial cells became flattened. On the other hand, basophilic tubules with peritubular lymphoid cell infiltration were multifocally observed in the cortex. In conclusion, PCP induced multiple renal cysts that developed in the collecting ducts of the outer medulla in neonatal rats, and it is suggested that epithelial cell proliferation may play some roles in the induction of cystic lesions.     

日粮硒水平对北京鸭生长及发病死亡的影响

将1日龄北京雏鸭180只按体重随机分成6组,每组30只。在低硒日粮中补充亚硒酸钠,调正各组日粮含硒水平为0.01,0.03,0.05,0.10,0.30及0.50ppm,测定体重,组织硒含量及血液谷胱甘肽过氧化物酶活性,研究日粮不同硒水平对北京鸭生长及发病死亡的影响。结果表明,日粮含硒在0.03ppm以下时,北京鸭死亡率高,增重差,缺硒病变严重。日粮含硒在0.05ppm以上才能维持北京鸭不发生缺硒死亡,其中以0.30ppm对生长发育更为适宜。     

The effect of lithium chloride on renal structure and sodium-potassium-adenosine triphosphatase activity in dogs

Lithium chloride was given intraperitoneally to dogs at a dosage of 125 mg/kg body weight for three days. Kidneys were removed for morphologic examination and quantitation of sodium-potassium-adenosine triphosphatase (Na-K-ATPase) activities in cortical and medullary tissue. Light microscopy showed no changes in the kidneys, but cytoplasmic vacuolation and dilatation of the cisternae of the endoplasmic reticulum were seen ultrastructurally in the epithelial cells of the distal tubule and cortical and medullary collecting ducts. Mean cortical Na-K-ATPase activity was 1.49 +/- 0.25 and 1.70 +/- 0.31 mumoles inorganic phosphate/mg protein/hour in control and experimental groups respectively. Mean medullary Na-K-ATPase activity was 4.71 +/- 0.41 and 5.01 +/- 0.47 mumoles inorganic phosphate/mg protein/hour in control and experimental groups respectively. It was concluded that lithium produced morphologic changes in the distal nephron, but had no effect on renal Na-K-ATPase activity.     

犊牛磺胺中毒的病理形态学观察

为了解犊牛磺胺中毒的病理形态学变化特点,对2头因磺胺中毒而死亡的犊牛进行了详细的病理学研究.结果表明,犊牛磺胺中毒的病理性损伤主要发生于肾脏和肝脏.宏观检查可见,肾脏肿胀,色泽变淡,皮质增宽,肾盂有多量豆腐渣样物质.肝脏肿胀,淡橘红色,质地脆易碎.微观检查可见,近曲小管上皮细胞肿胀,核浓缩,染色不良,大量上皮细胞脱落....     

东北虎肾组织形态结构的研究

通过对一只 8月龄东北虎 (雌 )肾脏组织形态结构的观察 ,结果表明 ,其肾小体体积较小 ,数量较少 ,且呈集中分布 ;在肾小体周围的远曲小管和近曲小管细小而稀少 ,并有较多的大血管分布 ;且近曲小管内微绒毛和侧突较稀疏 ,肾小囊腔较窄 ;髓放线、髓质部内的肾集合管细小而稀少 ,肾集合管及髓袢呈较疏松分布。观察结果还表明 ,东北虎肾脏的组织形态学结构有着显著的生态适应特征。     

Canine renal failure syndrome in three dogs

Three dead dogs were brought to the College of Veterinary Medicine, Kyungpook National University for study. Clinically, all the dogs showed emaciation, anorexia, depression, hemorrhagic vomiting and diarrhea for 7~10 days before death. All the clinical signs were first noted for about one month after feeding the dogs with commercial diets. At necropsy, all 3 dogs had severe renal damage with the same green-yellowish colored nephroliths in the renal pelvis. They also showed systemic hemorrhage and calcification of several organs, which might have been induced by uremia. Microscopically, necrosis, calcification and calculi were detected in the renal tubules, and especially in the proximal convoluted tubules and collecting ducts of the kidney. These findings were supportive of a mycotoxic effect, and especially on their kidneys. However, the precise cause of the toxic effect in these cases of canine renal failure could not be determined.     

Tubular reabsorption of protein by porcine kidneys during neonatal alimentary proteinuria

The ability of renal proximal tubular cells to reabsorb protein early in postnatal life was investigated using goat haemoglobin as tracer. The haemoglobin was intracardially administered to newborn piglets. The kidneys were fixed for light and electron microscopy 4 h later. Piglets killed immediately after birth and those allowed to suck colostrum for 4 h were used as controls. The proximal tubular cells of newborn, unsuckled piglets already contained absorptive vacuoles. Haemoglobin was absorbed in some absorptive vacuoles of the proximal tubules. The tracer was also demonstrable in the urine, mainly in the form of methaemoglobin. Although proximal tubular cells of newborn piglets are able to absorb protein, this absorption is of limited extent and excess protein is voided with the urine.     

Ultrastructural postmortem changes in chicken kidneys at 27 C

Ultrastructural postmortem changes of glomeruli, proximal tubules (PT), distal tubules (DT), and cortical collecting ducts (CD) were studied in adult chickens held at 27 C for 0, 1, 5, 10, 30, 60, or 360 minutes. There were marked ultrastructural alterations in tissues considered normal by light microscopy. The earliest changes, evident in immediately fixed samples, consisted of apical cell swelling of PT and DT epithelium and formation of smaller projections from glomerular capillary endothelium. Swelling increased with time, and gaps in cell membranes of tubular cytoplasmic blebs were evident by 10 minutes postmortem. The urinary space also contained cellular debris in immediately fixed samples and was filled by swollen epithelial cells by 10 minutes postmortem. Organelle changes were pronounced by 5 minutes in PT epithelium and by 10 minutes in DT epithelium. These initial changes consisted of dilation of mitochondrial and endoplasmic reticulum. Flocculent mitochondrial matrix densities were present by 5 minutes in PT epithelium and enlarged and appeared in other tubular segments as the time postmortem increased. Nuclear chromatin margination occurred first in 10-minute samples of PT and DT, and it was present by 30 minutes in glomerular epithelial cells. Chromatin clumping was present in PT and DT epithelium at 30 minutes and was followed by pyknosis and karyorrhexis at 360 minutes. These latter nuclear changes were not found in glomerular cells during the study. Nuclei of CD showed little change during the periods studied. The glomerular basement membrane appeared as a single hyaline membrane by 30 minutes postmortem, but this membrane and podocyte foot processes showed little further change by 360 minutes postmortem.     

Outbreaks of renal failure associated with melamine and cyanuric acid in dogs and cats in 2004 and 2007.

Sixteen animals affected in 2 outbreaks of pet food-associated renal failure (2 dogs in 2004; 10 cats and 4 dogs in 2007) were evaluated for histopathologic, toxicologic, and clinicopathologic changes. All 16 animals had clinical and laboratory evidence of uremia, including anorexia, vomiting, lethargy, polyuria, azotemia, and hyperphosphatemia. Where measured, serum hepatic enzyme concentrations were normal in animals from both outbreaks. All animals died or were euthanized because of severe uremia. Distal tubular lesions were present in all 16 animals, and unique polarizable crystals with striations were present in distal tubules or collecting ducts in all animals. The proximal tubules were largely unaffected. Crystals and histologic appearance were identical in both outbreaks. A chronic pattern of histologic change, characterized by interstitial fibrosis and inflammation, was observed in some affected animals. Melamine and cyanuric acid were present in renal tissue from both outbreaks. These results indicate that the pet food-associated renal failure outbreaks in 2004 and 2007 share identical clinical, histologic, and toxicologic findings, providing compelling evidence that they share the same causation.     

鸡实验性尿酸盐沉积症的肾脏病理学研究

将１２０只３５日龄迪卡商品代蛋用雏鸡随机分为高钙组、高蛋白组和对照组,分别饲喂高钙（含钙３．９％）、高蛋白（含粗蛋白４４．０％）和全价（含钙０．８％,粗蛋白１８．２％）饲料。试验第２５天,各组剖检５只鸡,作肾脏的光镜和电镜观察。结果表明,高钙、高蛋白饲料均能引发尿酸盐沉积。高钙组鸡肾脏肿大,表面及切面有大量尿酸盐结晶;输尿管扩张,管内充满白色尿酸盐沉积物。光镜观察,肾小球肿胀,近曲及远曲小管上皮细胞肿胀、变性。电镜观察,近曲小管上皮绒毛短缩、肿胀、断裂、脱落,线粒体肿胀、嵴断裂;远曲小管线粒体肿胀,呈现空泡,核呈锯齿型。高蛋白组眼观病变不明显,肾脏的光镜、电镜病变与高钙组相似,但损伤程度较轻。作者认为,高钙、高蛋白所致肾脏损害是鸡尿酸盐沉积症的重要原因。     

Prenatal Metanephrogenesis of The Camel: Morphological Evidence of Epithelial–Mesenchymal Interaction

The present investigation examined histogenesis of epithelial, stromal and angiogenic elements of the prenatal camel permanent or metanephric kidney. The primitive metanephros was first observed at the 13‐mm crown vertebral rump length (CVRL) stage as an ovoid structure composed of a centrally located epithelial ureteric bud and peripheral circumscribed masses of undifferentiated mesenchymal cells. The first morphological evidence of glomerulogenesis was observed at the 28‐mm CVRL stage. Developing renal corpuscles became obvious at the 35‐mm CVRL stage. At the 60‐mm CVRL stage, the epithelial renal pelvis gave rise to tubular branches that extended towards the cortical zone. These branches represented the presumptive collecting ducts. Differentiation of renal tubules into the proximal and distal convoluted tubules was observed at the 95‐mm CVRL stage. At the 130‐mm CVRL stage, the renal medulla was clearly delineated into medullary pyramids, which in association with the corresponding cortical caps formed the morphological basis of the renal lobar formation. A gradual nephrogenic decline was noticed from the 940‐mm CVRL on; however, the process of nephrogenesis persisted throughout all the studied foetal stages.     

复合酶制剂对北京填鸭胴体性状、体脂沉积和营养物质表观消化率的影响

本试验旨在研究饲粮复合酶制剂水平对北京填鸭胴体性状、体脂沉积和营养物质表观消化率的影响。选取35日龄健康、大小均匀的雄性北京鸭96只,随机分成4个组,每个组6个重复,每个重复4只鸭。对照组饲喂玉米-豆粕型基础饲粮,试验1组、试验2组和试验3组饲粮分别在基础饲粮中添加100、200和300 mg/kg的复合酶制剂,试验鸭在35和36日龄饲粮填饲量分别为260和300 g,之后5 d各组饲粮填饲量均为400 g/d。试验期为7 d。结果表明:1)饲粮复合酶制剂水平对北京填鸭屠体重、屠体率、全净膛重、全净膛率、腿肌重、腿肌率、龙骨长和胸宽均无显著影响(P0.05),而对北京填鸭增重、胸肌重、胸肌率、胸肌厚、肌胃重和肌胃率有显著影响(P0.05)。试验2组和试验3组北京填鸭胸肌重和肌胃重分别比对照组显著提高了7.88%和7.60%(P0.05)。2)饲粮复合酶制剂水平对北京填鸭皮脂重、皮脂率、腹脂重、腹脂率、肝脏重、肝重率和肝脂率均无显著影响(P0.05)。试验1组皮脂厚与对照组相比显著提高了16.2%(P0.05)。3)饲粮复合酶制剂水平对北京填鸭干物质和总能表观消化率无显著影响(P0.05),而对北京填鸭粗蛋白质表观消化率有显著影响(P0.05)。由此可见,饲粮中添加复合酶制剂对北京填鸭体脂沉积无显著影响,但能够提高饲粮中粗蛋白质表观消化率,并促进胸肌和肌胃的生长发育。