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  • Author or Editor: Christian M. Leutenegger x
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Abstract

Objective—To determine gene expression of selected molecular markers (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, IL-8, IL-10, procalcitonin [PCT], and transforming growth factor [TGF]-β) in the blood of healthy and sick foals.

Animals—28 sick foals without sepsis, 21 foals with sepsis, and 21 healthy foals.

Procedures—Total RNA was extracted from blood samples and converted into complementary DNA (cDNA). Gene expression was measured for the molecular markers by use of real-time PCR assay, and final quantitation was performed with the comparative threshold cycle method.

Results—Samples from all foals yielded transcription for all markers. Expression of TNF-α and TGF-β was significantly lower and that of IL-8 significantly greater in the sick-nonseptic and septic groups, compared with the healthy group. No significant difference in expression of IL-1β, IL-6, and PCT was found between the healthy group and the 2 sick groups. Expression of IL-10 was significantly greater in nonsurvivors, compared with survivors.

Conclusions and Clinical Relevance—The cytokine profile in foals with sepsis may suggest an immunosuppressive state. Expression of IL-10 may be a marker for identification of foals with a guarded prognosis.

Full access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

Abstract

Objective—To characterize isolates of Corynebacterium pseudotuberculosis from horses, cattle, and sheep in Colorado, Kentucky, Utah, and California in samples collected during perceived epidemics of infection (increased numbers of cases identified) in 2002 and 2003, and determine how closely isolates were related and their possible source.

Sample Population—54 isolates of C pseudotuberculosis from 49 horses, 4 cattle, and 1 sheep.

Procedures—Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) assay, PCR assay for the gene encoding the phospholipase D (PLD) toxin, biochemical analyses, and tests for susceptibility to 17 antimicrobial drugs were performed.

Results—All isolates reduced nitrate to nitrite, most yielded positive results for the PLD toxin gene, and all were susceptible to antimicrobial drugs. Ten genetic types were detected by use of RAPD PCR assay; types III to X were isolated from horses, cattle, or both in 1 or more states. Types III and IX were isolated from both horses and cattle. Types VII and VIII were isolated in only 1 state, but the number of isolates in these groups was small. In contrast, all other types were isolated in 2 or more states. All isolates from Utah were type III, but the other 3 states had isolates from more than 1 type.

Conclusions and Clinical Relevance—These data are consistent with a clonally expanding epidemic of infection in Utah and an increase in number of infections caused by multiple strains of C pseudotuberculosis not derived from a single source in the other states. The increase in number of infections could be the result of reporting bias, environmental factors facilitating infection, or host factors such as greater herd susceptibility. (Am J Vet Res 2004;65:1734–1737)

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in American Journal of Veterinary Research

Abstract

Objective—To determine whether 14-day topical ocular administration of high doses of feline recombinant interferon omega (FelFN) or human recombinant interferon alpha-2b (HulFN) solution improves clinical disease and decreases virus shedding in cats with naturally acquired viral keratoconjunctivitis.

Animals—36 cats with upper respiratory tract disease and ocular involvement.

Procedures—Cats received 1 drop of FelFN solution (1 × 106 U/mL), HulFN solution (1 × 106 U/mL), or saline (0.9% NaCl) solution (12 cats/group) in each eye twice daily for 14 days (beginning day 1). Oropharyngeal and conjunctival swab samples were collected from each cat before (day 0) and on day 14 of treatment for virus isolation (VI) and real-time quantitative PCR (RT-qPCR) testing to detect feline herpesvirus-1 and feline calicivirus. Subjective clinical scores were recorded on days 0, 3, 7, 10, and 14.

Results—The number of cats for which feline herpesvirus-1 was detected via VI or RT-qPCR assay was generally (albeit not always significantly) lower on day 14, compared with day 0 findings; however, findings on days 0 or 14 did not differ among groups. The number of cats for which feline calicivirus was detected via VI or RT-qPCR assay did not differ significantly between days 0 and 14 for any group. Clinical scores significantly decreased over the 14-day period but did not differ among groups.

Conclusions and Clinical Relevance—In cats with naturally occurring viral keratoconjunctivitis, bilateral ocular administration of high doses of FelFN or HulFN twice daily for 14 days did not improve clinical disease or virus shedding, compared with treatment with saline solution.

Full access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine the frequency of enteropathogens in dogs entering an animal shelter with normal feces or diarrhea.

Design—Cross-sectional study.

Animals—100 dogs evaluated at an open-admission municipal animal shelter in Florida.

Procedures—Fecal samples were collected within 24 hours after admission from 50 dogs with normal feces and 50 dogs with diarrhea. Feces were tested by fecal flotation, antigen testing, PCR assay, and electron microscopy for selected enteropathogens.

Results—13 enteropathogens were identified. Dogs with diarrhea were significantly more likely to be infected with ≥ 1 enteropathogens (96%) than were dogs with normal feces (78%). Only Clostridium perfringens enterotoxin A gene was significantly more common in dogs with diarrhea (64%) than in dogs with normal feces (40%). Other enteropathogens identified in dogs with and without diarrhea included hookworms (58% and 48%, respectively), Giardia spp (22% and 16%, respectively), canine enteric coronavirus (2% and 18%, respectively), whipworms (12% and 8%, respectively), Cryptosporidium spp (12% and 2%, respectively), ascarids (8% and 8%, respectively), Salmonella spp (2% and 6%, respectively), Cystoisospora spp (2% and 4%, respectively), canine distemper virus (8% and 0%, respectively), Dipylidium caninum (2% and 2%, respectively), canine parvovirus (2% and 2%, respectively), and rotavirus (2% and 0%, respectively).

Conclusions and Clinical Relevance—Dogs entered the shelter with a variety of enteropathogens, many of which are pathogenic or zoonotic. Most infections were not associated with diarrhea or any specific dog characteristics, making it difficult to predict the risk of nfection for individual animals. Guidelines for preventive measures and empirical treatments that are logistically and financially feasible for use in shelters should be developed for control of the most common and important enteropathogens.

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine the frequency of enteropathogens in cats entering an animal shelter with normal feces or diarrhea.

Design—Cross-sectional study.

Animals—100 cats evaluated at an open-admission municipal animal shelter in Florida.

Procedures—Fecal samples collected within 24 hours after admission from 50 cats with normal feces and 50 cats with diarrhea were tested by fecal flotation, antigen testing, PCR assay, and electron microscopy for selected enteropathogens.

Results—12 enteropathogens were identified. Cats with diarrhea were no more likely to be infected with ≥ 1 (84%) enteropathogens than were cats with normal feces (84%). Only feline coronavirus was significantly more prevalent in cats with diarrhea (58%) than in cats with normal feces (36%). Other enteropathogens identified in cats with and without diarrhea included Clostridium perfringens enterotoxin A (42% and 50%, respectively), Cryptosporidium spp (10% and 20%, respectively), Giardia spp (20% and 8%, respectively), Cystoisospora spp (14% and 10%, respectively), hookworms (10% and 18%, respectively), ascarids (6% and 16%, respectively), Salmonella spp (6% and 4%, respectively), astrovirus (8% and 2%, respectively), feline panleukopenia virus (4% and 4%, respectively), calicivirus (0% and 2%, respectively), and Spirometra spp (0% and 2%, respectively).

Conclusions and Clinical Relevance—In the present study, cats entered the shelter with a variety of enteropathogens, many of which are pathogenic or zoonotic. Most infections were not associated with diarrhea or any specific risk factors such as signalment, source, or body condition, making it difficult to predict which cats were most likely to be infected. It is not possible to test all shelter cats for all possible infections, so practical guidelines should be developed to treat routinely for the most common and important enteropathogens.

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine gene transcription for cytokines in nucleated cells in CSF of horses without neurologic signs or with cervical stenotic myelopathy (CSM), West Nile virus (WNV) encephalitis, equine protozoal myeloencephalitis (EPM), or spinal cord trauma.

Animals—41 horses (no neurologic signs [n = 12], CSM [8], WNV encephalitis [9], EPM [6], and spinal cord trauma [6]).

Procedures—Total RNA was extracted from nucleated cells and converted into cDNA. Gene expression was measured by use of real-time PCR assay and final quantitation via the comparative threshold cycle method.

Results—Cytokine genes expressed by nucleated cells of horses without neurologic signs comprised a balance between proinflammatory tumor necrosis factor-α (TNF-α), anti-inflammatory cytokines (interleukin [IL]-10 and transforming growth factor [TGF]-β), and Th1 mediators (interferon [IFN]-γ). Cells of horses with CSM mainly expressed genes for TNF-α, TGF-β, and IL-10. Cells of horses with WNV encephalitis mainly expressed genes for IL-6 and TGF-β. Cells of horses with EPM mainly had expression of genes for IL-6, IL-8, IL-10, TNF-α, IFN-γ, and TGF-β. Cells from horses with spinal cord trauma had expression mainly for IL-6; IFN-γ; TGF-β; and less frequently, IL-2, IL-10, and TNF-α. Interleukin-8 gene expression was only detected in CSF of horses with infectious diseases.

Conclusions and Clinical Relevance—Despite the small number of CSF samples for each group, results suggest distinct gene signatures expressed by nucleated cells in the CSF of horses without neurologic signs versus horses with inflammatory or traumatic neurologic disorders.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To estimate the analytic sensitivity of microscopic detection of Toxoplasma gondii oocysts and the environmental loading of T gondii oocysts on the basis of prevalence of shedding by owned and unowned cats.

Design—Cross-sectional survey.

Sample Population—326 fecal samples from cats.

Procedures—Fecal samples were collected from cat shelters, veterinary clinics, cat-owning households, and outdoor locations and tested via ZnSO4 fecal flotation.

Results—Only 3 (0.9%) samples of feces from 326 cats in the Morro Bay area of California contained T gondii–like oocysts. On the basis of the estimated tonnage of cat feces deposited outdoors in this area, the annual burden in the environment was estimated to be 94 to 4,671 oocysts/m2 (9 to 434 oocysts/ft2).

Conclusions and Clinical Relevance—Despite the low prevalence and short duration of T gondii oocyst shedding by cats detected in the present and former surveys, the sheer numbers of oocysts shed by cats during initial infection could lead to substantial environmental contamination. Veterinarians may wish to make cat owners aware of the potential threats to human and wildlife health posed by cats permitted to defecate outdoors.

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To correlate gene transcription of cytokines and chemokines with histologic inflammation in nasal biopsy specimens of cats.

Animals—25 study cats and 4 specific pathogen–free cats.

Procedure—One nasal biopsy specimen from each cat was submitted for routine histologic evaluation; a second was submitted for evaluation by use of a quantitative real-time polymerase chain reaction analysis with a fluorogenic probe (ie, TaqMan) for detection of cytokines and chemokines (interleukin [IL]-4, IL-5, IL-6, IL-10, IL-12 p40, IL-16, IL-18, interferon [IFN]-γ, tumor necrosis factor [TNF]-α, and the regulated on activation normal T cell expressed and secreted [RANTES] protein). Specimens were grouped histologically by degree of inflammation (none, mild, moderate, or severe). Linearized TaqMan signals for each gene were compared among histologic groups.

Results—Nasal biopsy specimens from specific pathogen–free cats were histologically normal, and cytokine transcription was low in these samples. As nasal inflammation in study cats worsened from absent (n = 3) to mild (4) to moderate (8) or severe (10), progressively and significantly increasing transcription of IL-6, IL-10, IL-12 p40, IFN-γ, TNF-α, and the RANTES protein was detected. Transcription of IL-4, IL- 5, IL-16, and IL-18 did not correlate with worsened histologic inflammation.

Conclusions and Clinical Relevance—Transcription of specific cytokines and chemokines in nasal tissue of cats progressively increased with severity of histologic evidence of inflammation, and IL-6, IL-10, IL-12 p40, IFN-γ, TNF-α, and the RANTES protein were markers of inflammation. Our data suggest that the nasal cavity of cats is biased toward a Th1 cytokine profile that is augmented by inflammation. (Am J Vet Res 2005;66:996–1001)

Full access
in American Journal of Veterinary Research