Histological and ultrastructural studies of renal lesions in Babesia canis infected dogs treated with imidocarb

Histological and electron microscopic examinations of the kidneys of 8 dogs suffering from fatal, naturally acquired Babesia canis infection and nephropathy are presented. Seven animals were treated with imidocarb dipropionate on average 4.5 days prior to death. Severe anaemia was present only in 2 cases. Degenerative histological changes observed mostly in the proximal convoluted tubules included vacuolar-hydropic degeneration, necrosis and detachment of renal tubular epithelial (RTE) cells from the basement membrane. Necrotic debris occasionally formed acidophilic casts within the tubules. In some cases, necrosis of the whole tubule was observed. Haemoglobin casts in the tubules and haemoglobin droplets in RTE cells seldom appeared. No significant histological changes were seen in the glomeruli. Ultrastructural lesions in RTE cells included nuclear membrane hyperchromatosis, karyopyknosis, karyolysis, swelling or collapse of mitochondria with fragmentation of cristae and vacuolar-hydropic degeneration in the endoplasmic reticulum and microvilli. Nuclear oedema was also observed. Many RTE cells exhibiting necrosis collapsed. Vacuolar-hydropic degeneration and necrosis were also observed in the glomerular and interstitial capillary endothelium. The severe acute tubular necrosis described in this study is probably the result of hypoxic renal injury. Systemic hypotension leading to vasoconstriction in the kidneys might be the most important cause of renal hypoxia in B. canis infections, but anaemia may also contribute to inadequate oxygenation. Imidocarb should be applied with caution in patients with possible renal involvement until further data become available on its potential nephrotoxicity in dogs.     

Cytopathic effects of Moraxella bovis on cultured bovine neutrophils and corneal epithelial cells

The effects of Moraxella bovis on the morphologic features of purified bovine neutrophils and bovine corneal epithelial cells were examined, using transmission and scanning electron microscopy and light microscopy. Within 2 minutes after incubation of bovine neutrophils with living M bovis, electron microscopic cellular changes included vacuolation, swelling, and loss of microplicae. Most of the neutrophils were lysed by 10 minutes of incubation. Human neutrophils phagocytosed the M bovis and remained intact, even after 30 minutes of incubation with the bacteria. Living M bovis killed bovine corneal epithelial cells in vitro. Sterile filtrates prepared from 6-hour shaker cultures of M bovis also killed bovine corneal epithelial cells, but the cytotoxic activity was less than that produced by the living bacteria. Cellular changes were first observed in specimens collected 1 hour after corneal cell monolayers were inoculated with sterile culture filtrates. The changes in these cells included pit-like lesions on the cellular surface, cellular separation, and vacuolation.     

Morphologic alterations in the anterior lens capsule of canine eyes with cataracts

OBJECTIVE: To examine the morphologic changes in the anterior lens capsule and lens epithelium of canine eyes with cataracts. SAMPLE POPULATION: Anterior lens capsules from the eyes of 25 dogs with cataracts and from an additional 10 canine globes with lenses subjectively assessed to be normal. PROCEDURE: Thickness of each anterior lens capsule was measured by use of a digital microscopic camera and imaging software. All 25 capsules from eyes with cataracts were submitted for light microscopy; 4 were also submitted for electron microscopy. RESULTS: Thickness of the anterior lens capsule increased with age for the normal lenses and the lenses with cataracts; the change with age was similar for both groups. Light microscopy revealed fibrous metaplasia of lens epithelial cells in 7 of 25 anterior lens capsules with focal thickenings of the posterior aspect of the capsule. Electron microscopy revealed deposition of collagen and basement membrane-like material by fibroblast-like cells. CONCLUSIONS: Results indicate that thickness of the anterior lens capsule in dogs increases with age and that this increase in thickness is not significantly different between normal lenses and lenses with cataracts. In addition, epithelial cells from lenses with cataracts may undergo metaplasia to form plaques composed of fibrous tissue and ectopic basement membrane produced by epithelial cells.     

Clinical, histologic, and histochemical study of imidocarb diproprionate toxicosis in goats.

The toxic effects of imidocarb diproprionate (3,3'-bis [2 imidazolin-2yl]-carbanilde diproprionate) were evaluated in adult goats given (intramuscular injection) a lethal dose (6.75 mg/kg). The immediate clinical signs of toxicosis were transient excessive salivation and diarrhea. Anorexia, dyspnea, recumbency, and death occurred between postinjection days (PID) 4 and 8, during which time 7 goats died and 4 moribund goats were euthanatized. There were marked increases in mean serum urea nitrogen concentration and significant increases in serum glutamic oxalacetic transminase activity and in the mean number of circulating neutrophils after PID 4. Renal hyperemia and enlargement were evident by PID1. Serosanguineous fluid in the trachea and major bronchi, pulmonary congestion and edema, hydrothorax, hydroperitoneum, and less frequently hydropericardium were observed on and after day 4. Microscopic renal tubular lesions rapidly progressed from pyknotic epithelial nuclei observed at 6 and 12 hours to acute tubular necrosis of epithelium of the proximal convoluted tubules on days 1 and 2. Pulmonary congestion and edema; hemorrhage into alveoli, bronchioles, and bronchi; and intracytoplasmic lipid vacuoles within the hepatocytes in the periacinar zones of the hepatic lobules were observed on or after day 4. Succinic dehydrogenase and adenosine triphosphatase activities decreased progressively in the epithelial cells of the proximal convoluted tubules. The decreases in cellular enzymatic activity occurred shortly after the appearance of microscopic lesions in the tubular epithelium.     

A scanning and transmission electron microscopic study of rotavirus-induced intestinal lesions in neonatal gnotobiotic dogs

Neonatal gnotobiotic dogs orally inoculated with canine rotavirus had ultrastructural changes limited to the jejunal and ileal regions of the small intestine. Early scanning electron microscopic findings consisted of swollen villus epithelial cells, denuded foci on intestinal villi, and slight to moderate villus atrophy. Later changes were slight villus atrophy with no denuded intestinal villi. Transmission electron microscopic changes in villus epithelial cells from 12 to 48 hours post-inoculation included: rotavirus particles associated with intracytoplasmic vacuoles near the terminal web and apical tubules; viral particles in dilated cisternae of rough endoplasmic reticulum; and moderate numbers of necrotic cells having no microvilli, swollen mitochondria, membrane-bound lipid-like material in the cytoplasm, clumped chromatin around the periphery of the nucleus, and disruption of the cytoplasmic membrane. In jejunum and ileum at 72 to 154 hours post-inoculation, there were fewer necrotic villus epithelial cells and fewer virus particles. In addition, the ultrastructural morphology of the majority of the villus epithelial cells was similar to crypt epithelium. These studies showed that rotavirus infected the villus epithelial cells with subsequent propagation of the rotavirus and destruction of villus epithelial cells.     

Morphological study of the effects of the GnRH superagonist deslorelin on the canine testis and prostate gland

The present study is part of a programme of research designed to evaluate the efficacy of the GnRH superagonist,deslorelin (D-Trp6-Pro9-des-Gly10-LHRH ethylamide), as a contraceptive for male dogs. Adult dogs were assigned to a completely randomized design comprising six groups of four animals. Each dog in the control group received a blank implant (placebo) and each dog in the other five groups received a 6 mg deslorelin implant. One group of deslorelin treated dogs was sacrificed on each of days 16, 26, 41, 101 and 620, and testicular and prostate tissues were collected for study by light and electron microscopy. On days 16 and 26 after implantation, we observed partial disruption of the seminiferous tubules, with early spermatids shed into the lumen. On days 41 and 101 after implantation, 90–100% of the seminiferous tubules were atrophic and aspermatogenic.On day 101 after implantation, 99% of all sections showed atrophy of the epithelium and shrinkage of epithelial height in the ductus epididymides. On days 41 and 101 after implantation, prostate tissue showed complete atrophy of the glandular epithelium (100% of sections) and an apparent increase in the relative proportion of connective tissue. At the electron microscopic level, in dogs treated with deslorelin for 41 and 101 days, the Sertoli cells were smaller and their nucleoli appeared smaller than in the control dogs. The nucleoli of the Leydig cells were atrophied and prostate glandular epithelium showed reduced epithelial height, a trophy of the nucleolus and an absence of secretory granules.Tissues collected during the recovery phase revealed a complete recovery of spermatogenesis. In conclusion, slow release implants containing deslorelin induce a striking a trophy of the testes and prostate gland by 26 days after implantation, explaining the previously reported loss of ejaculate and arrest of sperm output. At histological level,the entire process appears to be completely reversible, in accordance with data on endocrine variables and semen production.     

Transmissible gastroenteritis in neonatal dogs: experimental intestinal infection with transmissible gastroenteritis virus.

Fourteen neonatal dogs (4 through 11 days of age) were exposed orally to the Purdue strain of transmissible gastroenteritis (TGE) virus, and six dogs of similar age were noninoculated controls. Clinical signs of enteric disease did not develop. Both exposed and control dogs had normal fecal passages and appetite throughout the experiment. Jejunal epithelium from dogs euthanatized at 12, 24, 48, and 96 hours and at 10 days after exposure did not exhibit morphologic alterations detectable by light microscopy. Electron microscopic examination indicated that jejunal epithelial cells contained TGE viral particles as early as 12 hours after dogs were exposed. There were no apparent morphologic alterations or signs of desquamation of virus-infected cells, however. Results of pig transmission studies indicated that viable TGE virus was in jejunal tissue of the dogs as early as 12 hours and as late as 10 days after exposure to the virus.     

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.     

Distribution and amount of cathepsin B in gentamicin-induced acute kidney injury in rats

The aim of our study was to investigate how the distribution and amount of cathepsin B change during acute kidney injury. The research was done on a rat model of acute kidney injury that was induced by nephrotoxic antibiotic gentamicin. Gentamicin was injected at a dose of 40 mg/kg body weight (the first treated group) and 80 mg/kg body weight (the second treated group) for 14 days. Control groups received injections of physiological saline only. One day after the last injection, animals were euthanized, dissected and kidney samples were taken and fixed in 10% buffered formalin. Tissue sections were stained with haematoxylin and eosin, periodic Acid Schiff (PAS) and Oil-red-O. Immunohistochemistry was used for the demonstration of cathepsin B. Vacuolar degeneration of the proximal convoluted tubules was the most prominent pathologic lesion found in the first treated group, while necrosis prevailed in the second treated group in the same localisation. In both treated groups significantly weaker immunohistochemical reaction for cathepsin B was noticed in the proximal convoluted tubules in comparison to the control groups (P < 0.05). The decrease of positive reaction was the largest in the proximal convoluted tubules of the outer renal cortex. Stronger positive reaction for cathepsin B, although not statistically significant, was found in the proximal straight tubules (P > 0.05), as well. However, more numerous cathepsin B-positive large granules appeared in the proximal straight tubules of the second treated group then in the second control group (P < 0.05). We can conclude that the amount of cathepsin B in the affected proximal convoluted tubules significantly decreases along the increased severity of the histopathological lesions of the proximal convoluted tubuls, the amount of enzyme in the well preserved proximal straight tubules increases and more cathepsin B-positive large granules appear in the cytoplasm.     

The effects of microwave-irradiated fixation for postmortem changes of the kidney

In the present study, we evaluated the advantages of microwave-irradiated fixation for postmortem autolysis of the kidney. Mouse kidneys, sampled at 0, 1, 3, 5, 10, 15, 20 and 25 hr after death, were fixed with 10% neutral formalin by microwave irradiation (MWI; 20 sec/500 W) and by conventional immersion. They were then examined with light and electron microscopy, morphometrics and immunohistochemicals. Light microscopic and morphometric observations showed that structural preservation effect of MWI was limited to the proximal convoluted tubules at 25 hr. Contrary, mild ultrastructural damage by MWI was found in the glomeruli at 0 and 15 hr. Immunohistochemistry for renin and alpha-smooth muscle actin showed no apparent differences between MWI and the immersion.     

Comparative morphology of esophageal mucosa and submucosa in dogs from 1 to 337 days of age

Scanning electron microscopy, light microscopy, and morphometric analyses were used to examine the morphology of the tunica mucosa-tela submucosa of the cervical, thoracic, and abdominal parts of the esophagus of healthy dogs from birth to 337 days of age from 3 litters of dogs whelped and reared under controlled conditions. Apart from the absence of a lamina muscularis mucosae from the cervical part of the esophagus of all dogs examined, little regional variation was detected. However, morphologic variation associated with age was observed. The number and complexity of microplicae on surface epithelial cells, as observed with the scanning electron microscope, increased with age, particularly between 1-day-old and 21-day-old dogs. Although scanning electron microscopy revealed typical duct openings from the submucosal glands in 1-day-old dogs, light microscopy revealed few functional glands. These glands gradually developed into a complex strata of tubules and acini during the time that secretory activity and the volume fraction of the tunica mucosa-tela submucosa occupied by glands increased with age. The presence of ciliated cells in the esophagus of 1-day-old dogs was apparent with scanning electron and light microscopes. Ciliated cells were always most abundant in the abdominal part of the esophagus. Finally, morphometric analyses revealed a significant increase in epithelial and connective tissue compartment thicknesses, as well as a marked increase in the volume fraction occupied by glands between 1 day and 161 days after dogs were whelped. A smaller decrease in all 3 measures occurred between 161 and 337 days after the dogs were whelped.     

Localisation and expression of TRPV6 in all intestinal segments and kidney of laying hens

1. The aim of this study was to investigate the localisation and expression of the epithelial Ca2+ channel TRPV6 (transient receptor potential vanilloid channel type 6) in different intestinal segments and kidney of laying hens during peak lay. 2. Immunohistochemical analysis of the intestine indicated that TRPV6 was localised to the brush-border membranes of the duodenum, jejunum, ileum, caecum, and rectum. Expression was weaker in the rectum, and little or no expression was found in crypt and goblet cells. In addition, TRPV6 mRNA was quantified amongst different intestinal segments, and expression was highest in the duodenum and jejunum. Furthermore, Western blotting indicated that the duodenum expressed the greatest amount of TRPV6 and the rectum the least with the other segments expressing intermediate levels. 3. In the kidney, distinct immunopositive staining for TRPV6 was detected at the apical domain of the distal convoluted tubules (DCT) and medullary connecting tubules (CNT). Interestingly, distribution of TRPV6 extended to the proximal convoluted tubules (PCT). Furthermore, the kidney expressed lower TRPV6 mRNA and protein levels compared with that in the duodenum. 4. In conclusion, the epithelial Ca2+ channel TRPV6 is strongly expressed in the apical cells of the entire intestine and the renal tubules, suggesting that active Ca2+ transcellular transport plays a crucial role in dietary calcium (re)absorption in laying hens.     

Studies on the pathogenesis of heart lesions in dogs infected with pseudorabies virus.

Pseudorabies virus was inoculated by various routes into dogs to determine the relationship of pseudorabies virus to the development of heart lesions. Electrocardiograms and serum samples for lactate dehydrogenase isoenzymes were taken twice daily. Transitory and persistent arrhythmias were a consistent finding. Heart lesions were noted within each of the inoculated groups. These changes varied from severe multifocal areas of hemorrhage and myocardial degeneration to small zones of myolysis. A ganglioneuritis of the stellate ganglia and autonomic ganglia within the heart were also consistent findings. Herpes-like viral particles were found by electron microscopy in various autonomic ganglia and in myocardial endothelial cells. No viral particles were found in myocardial cells. Significant increases in lactate dehydrogenase-1 were noted. It was concluded that fatal arrhythmias resulted from pseudorabies virus infections in the dog after the occurrence of myocardial and ganglionic lesions.     

Ultrastructural study of ochratoxicosis in quail (Coturnix coturnix japonica)

Ultrathin sections of liver and kidney from 11-week-old quail fed, from day-old, ochratoxin A (OA) (mixed in the diet at levels of 4 and 8 ppm) were examined in the electron microscope. Pathological changes in the kidneys were limited to the proximal convoluted tubules (PCT) and glomeruli. In the PCTs, abnormal mitochondria and excessive numbers of lipid droplets were the principal findings with glomeruli showing thickened basement membranes. Swollen mitochondria and variable glycogen deposition were the chief features present in the livers. It is suggested that OA is more hepatotoxic in quail than in broilers, as well as being nephrotoxic.     

Comparative ultrastructural study of normal, adenomatous, carcinomatous, and hyperplastic cells of canine hepatoid circumanal gland

Transmission electron and light microscopy studies were carried out on morphologic characteristics of cellular organelles of normal and different neoplastic cells of hepatoid circumanal glands of 16 dogs. The ultrastructural features of the reserve and transitional cells in the adenomas and carcinomas are highly suggestive of different carcinogenic stimuli for each form of hepatoid circumanal gland neoplasm. The reserve cells, in response to the carcinogenic stimuli, appear to be growing at the expense of the degenerating glandular cells.     

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.     

Renal dysfunction in dogs with pyometra

Renal function and pathologic changes in 27 dogs with pyometra were studied. Evaluation included CBC; serum biochemical evaluation; urinalysis; urine and uterine bacteriologic culture; uterine morphologic features; and light, electron, and immunofluorescent microscopic evaluation of renal tissues. Measurements of 24-hour creatinine clearance, protein excretion, Na excretion, and urine volume were made in 12 dogs without azotemia. Of 27 dogs, 26% were azotemic and 89% had a urine sp gr less than 1.035. Glomerular filtration rate was reduced in 75% of 12 dogs without azotemia. None of these 12 dogs was proteinuric. Examination of renal biopsy specimens revealed a high prevalence of mild tubulointerstitial nephritis, but few specific glomerular lesions. Minimal immunofluorescence was detected within the mesangium in 18% of the dogs. Immunofluorescence was not associated with the interstitium or tubules. Urinary tract infection was detected in 22% of the dogs. Escherichia coli and Klebsiella were recovered from the uterus in 59 and 15% of the dogs, respectively. Low urine specific gravity values were obtained from dogs without azotemia and from dogs with uterine cultures considered negative for E coli and other gram-negative bacteria. The reduction in glomerular filtration rate was a functional abnormality not correlated with structural damage in the glomerulus.     

Distribution and form of uranium-containing deposits in chickens treated with uranyl nitrate

The kidneys of chicks treated with uranyl nitrate were examined by electron microscopy. Most deposits of electron-dense material containing uranium were found in the lumina of distal tubules and collecting ducts of kidneys collected 12-24 hours post-treatment. Some deposits were present in extracellular spaces between adjacent cells. Only occasionally were deposits found intracellularly where they were associated with localized cellular degeneration. A generalized cellular degeneration over the whole kidney was seen 72-96 hours post-treatment, but this was not directly associated with the deposits of electron-dense material.     

Inflammation induced by subcutaneous turpentine inoculation of young American alligators (Alligator mississippiensis)

Turpentine-induced skin lesions in young American alligators (Alligator mississippiensis) kept at 25 C were used to study inflammatory response in a reptile. Skin harvested at intervals between 4 hours and 30 days after inoculations were done had no gross changes until days 24 to 26, when superficial skin necrosis was evident. Early responses of congestion and dermal edema (4 to 8 hours) were seen by light microscopy, and these were followed by necrosis and granulocyte migration (1 to 3 days). Later, there was predominance of monocytic cells, including vacuolated macrophages (7 to 30 days). Evident at 14 days and prominent by day 30 were central dermal zones of necrotic debris surrounded by orderly palisades of vacuolated multinucleated giant cells and capillary-laden immature fibrous connective tissue. Systemic illness or visceral lesions were not observed. Controls, given inoculations of sterile saline solution, had no gross or microscopic changes.