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  • Author or Editor: Christian M. Leutenegger x
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in Journal of the American Veterinary Medical Association

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

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 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 describe dogs with detected Ancylostoma caninum anthelmintic treatment resistance markers in Canada.

ANIMALS

11 client-owned dogs with fecal quantitative PCR (qPCR) assay detected A caninum with benzimidazole (BZ) resistance genotypic markers.

METHODS

Signalment, presenting concern, duration of clinical signs, fecal testing, treatment, and outcomes were obtained. Where available, follow-up data were collected via telephone or email with the primary veterinarian.

RESULTS

Ancylostoma spp was detected from 184/32,205 dog fecal samples by reference laboratory qPCR surveillance, between May 15, 2022, and April 26, 2023. 11 of these 184 samples had A caninum with genetic BZ F167Y resistance marker detection. 4 dogs had not traveled outside Canada, 6 had been imported from the US, and the travel history was unclear in 1 dog.

7 of the dogs had gastro-intestinal signs (diarrhea or soft stool) on initial presentation. Clinical improvement was reported in 6 of these dogs (resolution of diarrhea and soft stool), with 1 dog lost to follow-up. All 11 dogs received anthelmintic treatment (varied drugs and duration).

CLINICAL RELEVANCE

Identification of genetic markers of BZ resistance raises concerns about the potential animal and human impacts of resistant hookworms. 4 dogs lacked an origin from or travel history to the US, indicating true emergence and/or novel spread within Canada, not just importation from an area where resistance has been reported. Fecal surveillance was performed with a qPCR test incorporating treatment (BZ) resistance markers. There is a need to raise clinician awareness around treatment-resistant hookworm in dogs and the capability of fecal surveillance for genotypic and phenotypic resistance.

Open access
in American Journal of Veterinary Research

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.

Full access
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.

Full access
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)

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To describe the novel PCR diagnosis and outcome of intestinal Echinococcus multilocularis in a dog.

ANIMAL

A 13-month-old female intact dog with naturally occurring intestinal E multilocularis.

CLINICAL PRESENTATION, PROGRESSION, AND PROCEDURES

The 13-month-old dog initially presented with a reduced appetite and weight loss and then developed hematochezia. The clinical history included a lack of endoparasite preventive care (fecal testing, deworming), exposure to coyotes, fox, sheep, and rodents and the dog had intermittently been fed a raw food diet. Physical examination revealed a thin dog, with a 2/9 body condition score, that was otherwise clinically unremarkable. A fecal sample was submitted for screening for gastrointestinal parasites as part of an infectious disease assessment. The fecal PCR test reported detection of E multilocularis. This result was sequenced as the European haplotype E3/E4. Centrifugal flotation (same sample) did not detect taeniid eggs.

TREATMENT AND OUTCOME

The dog was treated with metronidazole, maropitant, and milbemycin oxime/praziquantel. Clinical improvement was noted within 48 hours. No DNA of E multilocularis was detected in a fecal sample collected approximately 10 days after treatment. The dog’s owner was advised to provide monthly deworming (praziquantel) for all dogs on the property and to contact their human health-care provider due to potential zoonotic exposure risk.

CLINICAL RELEVANCE

Increasing detection of E multilocularis is occurring in dogs in Canada and the US. Alveolar echinococcosis can cause severe disease in dogs and humans. Fecal PCR detection and surveillance may alert practitioners to canine intestinal cases and allow dogs to serve as sentinels for human exposure risk.

Open access
in Journal of the American Veterinary Medical Association