Comparison of viability and infectivity of Cryptosporidium parvum oocysts stored in potassium dichromate solution and chlorinated tap water

The present study was undertaken to compare the viability and infectivity of Cryptosporidium parvum oocysts that had been stored for 1, 4, 7, 10, 13, 16, 20, 25 and 30 months at 4 degrees C in 2.5% potassium dichromate (Cr) or chlorinated tap water, respectively. An excystation protocol was performed in vitro to evaluate viability. One hundred and eighty female BABL/c mice were used to evaluate the infectivity of oocysts by investigating the prepatent period of C. parvum infection, the quantity of oocysts excreted, and the number of parasites that colonized the villi of the ileum. The results showed that C. parvum oocysts preserved in Cr for 1-16 months or in water for 1-13 months were capable of excystation in vitro and infection of mice. The excystation rates of oocysts and the prepatent periods in mice infected by oocysts stored in Cr and water were not significantly different (p>0.05), and there was a strong correlation between prepatent period and duration of oocyst storage (Cr: R2=0.92; water: R2=0.98). There were no significant differences in oocyst shedding from feces or parasitism of the terminal ilea of mice by Cryptosporidia between the two storage media (p>0.05). In conclusion, C. parvum oocysts may be stored at 4 degrees C in water instead of Cr for the purposes of laboratory research. However, the presence of viable C. parvum oocysts in water is a severe challenge to the drinking water treatment industry.     

Scanning electron microscopy of Heinz bodies in feline erythrocytes.

Whole blood and partially lysed blood films from 5 cats having 20 to 91% of the erythrocytes containing Heinz bodies were examined, using the light microscope and the scanning electron microscope. Heinz bodies were detected in the intact erythrocytes from 3 of the cats as abruptly elevated and distinctly demarcated protuberances in various shapes, sizes, and locations. The Heinz bodies were located just beneath the cell membrane either centrally or near the cell margin and varied in their projectional magnitude. Brilliant cresyl blue staining of blood of these 3 cats revealed prominent Heinz bodies within, and projecting from, the erythrocytes. In contrast, Heinz bodies were not identified on scanning electron microscopy of intact erythrocytes of the remaining 2 cats even though Heinz bodies were found on their blood films stained with brilliant cresyl blue. Scanning electron microscopy of partially lysed blood smears of all 5 cats revealed Heinz bodies of various sizes in the erythrocyte ghosts. Furthermore, blood smears from the 3 cats having distinct Heinz bodies in intact erythrocytes revealed small dense intracellular granules distributed singly or coalesced in small clumps. Further aggregation of these clumps was assumed to result in the formation of a single large Heinz body. The 3-dimensional nature of Heinz bodies was clearly apparent in lysed blood smears.     

Susceptibility of canine, feline and human erythrocytes to oxidant-mediated injury

e relative susceptibilities of feline, canine and human erythrocytes to in vitro hydrogen peroxide-induced lipid peroxidation and hemolysis were studied. At 15 minutes following exposure to hydrogen peroxide, cat erythrocytes had higher concentrations of lipid peroxidation by-products and a greater percent hemolysis as compared to dogs and humans. Erythrocytes from cats with induced sterile abscesses had lower concentrations of the antioxidant glutathione, but they did not have detectable concentrations of lipid peroxides nor were they more susceptible to in vitro lipid peroxidation or hemolysis.     

Sperm viability in ram semen diluted and stored in three different extenders.

Semen was collected with an artificial vagina from 4 one-year-old rams, in order to study the changes in sperm motility and membrane integrity of spermatozoa split-diluted and stored at 5 degrees C during 7 days in sodium citrate, Tris, and milk-based extenders, respectively. Sperm motility was assessed subjectively and sperm membrane integrity was determined using the fluorescent probes Calcein-AM and Ethidium homodimer. Representative samples were studied using scanning electron microscopy (SEM). The average incidence of sperm motility decreased over time in all the extenders (p < 0.001). The incidence of spermatozoa showing progressive motility and intact plasma membrane was significantly higher in semen diluted with sodium citrate than in the other 2 extenders following 4 days of dilution until the end of the study. Evaluation with SEM confirmed the findings obtained with the supra vital fluorescent dyes. The results of the present study indicated that there were no differences between sodium citrate-, Tris- or milk-based extenders when ovine liquid semen was stored at 5 degrees C during a short period (2 days). However, when semen was stored for longer time, spermatozoa in the sodium citrate-based extender sustained its viability better.     

Ex vivo viability of canine and feline sarcomas: a pilot study

Assay-based chemotherapeutic protocols are common in human gynecologic oncology, most notably for patients with ovarian or breast cancer. The current study examines ex vivo incubation conditions necessary for the assessment of sarcomatous tumor response to potential chemotherapeutic drugs. Slices of sarcomatous tumors were incubated in one of two culture media. Viability indices were measured and compared across time and between media. Neither medium was sufficient to support the growth of sarcomatous tumor tissue slices based on the indices studied. It is likely that sarcomatous tumors require a different approach for ex vivo assessment than their epithelial counterparts. Our long-term goal is to incubate tumor slices with chemotherapeutic agents to predict the in vivo tumor response based on the maintenance or loss of slice viability within this system.     

The quality of cryopreserved sperm collected from feline caudal epididymides stored at room temperature

On the assumption that animals of wild feline species died in the field, caudal epididymal sperm were cryopreserved following storage of the feline epididymides at 20°C for 0-24 hr, and their qualities were observed. Compared to the qualities at 0 hr, no significant differences were noted following 12 hr of storage at 20°C. On comparison of the qualities between caudal sperm cryopreserved after 24 hr storage at 4°C and after 12 hr at 20°C followed by 12 hr storage at 4°C, no significant differences were noted. These findings suggest that the cryopreserved sperm collected from epididymides of dead animals might be useful for artificial insemination if cryopreservation was performed within 12 hr exposure to ambient temperature.     

Effect of a bioflavonoid dietary supplement on acetaminophen-induced oxidative injury to feline erythrocytes

OBJECTIVE: To determine the effect of a commercial bioflavonoid antioxidant on acetaminophen-induced oxidative injury to feline erythrocytes. DESIGN: Randomized controlled study. ANIMALS: 45 healthy age-matched cats. PROCEDURE: Cats were assigned to 3 experimental groups. Groups 1 and 3 received a bioflavonoid antioxidant (10 mg/d) orally for 2 weeks. Groups 2 and 3 received an oxidative challenge with acetaminophen (90 mg/kg [41 mg/lb] of body weight, PO) on day 7. Packed cell volume, percentage of erythrocytes with Heinz bodies, blood methemoglobin concentration, and blood reduced and oxidized glutathione concentrations were determined at various times during the 2-week study period. RESULTS: Adverse effects were not associated with bioflavonoid antioxidant administration alone. Acetaminophen administration resulted in a significant increase in methemoglobin concentration in groups 2 and 3; differences were not detected between these groups. Heinz body concentrations in groups 2 and 3 increased after acetaminophen administration; however, the increase in cats that received the antioxidant was significantly less than in group-2 cats. Total blood glutathione concentrations did not change significantly in groups 2 and 3 after acetaminophen administration; however, ratio of reduced to oxidized glutathione concentration increased significantly after administration in group-2 cats, compared with group-3 cats. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of bioflavonoid antioxidants to cats at risk for oxidative stress may have a beneficial effect on their ability to resist oxidative injury to erythrocytes.     

Endocytosis of erythrocytes in vivo and particulate substances in vitro by feline neoplastic mast cells.

Clinical evidence for the phagocytic capability of neoplastic feline mast cells was provided by recognition of endocytosed erythrocytes in seven of 12 cytological smears of mast cell neoplasms, particularly in those cells collected from splenic tumors. The capability of these neoplastic mast cells to endocytose particulate substances was also studied in vitro. Evidence is presented that under cultural conditions, feline neoplastic mast cells are capable of endocytosing a variety of substances including polystyrene latex microspheres, zymosan particles, horse spleen ferritin, salmon sperm nuclei, horseradish peroxidase, and carbon particles.     

Ocular effects of topical 0.03% bimatoprost solution in normotensive feline eyes

OBJECTIVE: The current study was undertaken to evaluate the effects of topically applied bimatoprost, an ocular hypotensive lipid, on intraocular pressure (IOP) and pupil size (PS) in healthy cats. ANIMAL STUDIED: Nine European Shorthair cats free from clinically relevant ocular abnormalities were used in the study. PROCEDURES: Pretreatment baseline measurements of IOP and PS were obtained bilaterally at 8 am, 2 pm, and 8 pm for five consecutive days (days 1 to 5). Then the cats received one drop twice daily (10 am and 6 pm) of bimatoprost ophthalmic solution 0.03% (Lumigantrade mark, Allergan Inc., Irvine, CA USA), in one randomly selected eye and one drop of artificial tears in the fellow eye (control eye) for 5 days (days 6 to 10). Values for IOP and PS were obtained under the same conditions as in the pretreatment phase. The potential for ocular irritation following bimatoprost application was also evaluated. RESULTS: During the pretreatment period, the mean IOP and mean PS were not significantly different between the eyes subsequently treated with bimatoprost and those subsequently determined as controls. During the treatment period, the mean IOP in bimatoprost-treated eyes was not significantly lower than in control eyes (14.2+/-2.3 vs. 14.5+/-2.8 mmHg). Mean IOP in control eyes was not significantly changed at any time during the study period. A marked reduction of PS was seen in all bimatoprost-treated eyes, but no other clinically relevant side effects were observed. CONCLUSION: Twice daily topical applications of bimatoprost produced miosis but had no significant effect on IOP in healthy cats.     

Influence of phosphate-buffered sucrose solution on early graft function in feline renal autotransplantation

Graft perfusion with cold heparinized saline has known to induce ischemia and reperfusion injury in feline kidney transplantation. In this study, the effects of phosphate-buffered sucrose solution and heparinized saline solution on early kidney graft function were compared in feline kidney autotransplantation. Perfusion of grafts with or without hypothermic storage with chilled phosphate-buffered sucrose solution prevented ischemia and reperfusion injury despite a very short ischemic time. The results of our study suggest that phosphate-buffered sucrose perfusion and storage solution should be effective to reduce ischemia and reperfusion injury despite a very short ischemic time in feline kidney transplantation.     

Preservation of feline anti-Toxoplasma gondii antibody activity using blood absorbed on filter paper stored under different conditions

The present study deals with the successful long-term preservation of feline anti-Toxoplasma gondii antibody activity on filter paper strips stored with silica gel. When filter paper strips were kept at 25 degrees C with silica gel, antibody activity in blood samples was preserved for at least six months.     

Prevalence of feline leukaemia provirus DNA in feline lymphomas

A significant drop in the prevalence of feline leukaemia virus (FeLV) antigenaemic cats and antigen-associated lymphomas has been observed after the introduction of FeLV vaccination and antigen-testing with removal of persistently antigenaemic cats. However, recent reports have indicated that regressively infected cats may contain FeLV provirus DNA and that lymphoma development may be associated with the presence of provirus alone. In the present study, we investigated the presence of FeLV antigen and provirus DNA in 50 lymphomas by immunohistochemistry and semi-nested polymerase chain reaction, respectively. Interestingly, almost 80% of T-cell lymphomas and 60% of B-cell lymphomas contained provirus DNA while only 21% of T-cell lymphomas and 11% of B-cell lymphomas expressed FeLV antigen. In conclusion, our results support previous hypotheses that vaccination and removal of persistently antigenaemic cats have led to a drop in FeLV antigen-expressing lymphomas. However, FeLV provirus DNA is still present in a high percentage of feline lymphomas.