Canine ocular onchocerciasis in New Mexico: Risk factors for disease.

OBJECTIVE: To determine whether dogs are at variable risk of developing canine ocular onchocerciasis based on coat color or size, factors that may influence feeding behavior of the putative vectors of Onchocerca lupi¸ the filarial nematode parasite causing this disease. ANIMALS STUDIED: One hundred twenty-five client-owned dogs diagnosed with confirmed or suspected onchocerciasis. 1255 dogs without signs of this disease were utilized for comparison. PROCEDURES: Dogs lacking signs of canine onchocerciasis were assessed for coat color and weight. Proportions of dogs with these characteristics were used to predict signalment in a group of dogs with this disease, if the investigated characteristics were unrelated to disease risk. Predicted values were compared statistically with observed values in the diseased dog population. Black fly color preference was assessed utilizing black, brown, and white traps and statistically assessing any differences in trapping based on trap color. RESULTS: Results suggest that large, black dogs are more likely to develop canine ocular onchocerciasis than was predicted by chance alone (p = 0.012). Results for smaller dogs with other coat colors were not significant. An increased risk for black dogs was supported by trapping data, as black flies were trapped significantly more often in black traps (p < 0.001). CONCLUSION: While factors other than size and color probably also influence the likelihood of O. lupi infection and disease development, our results may assist clinicians in their diagnosis of canine ocular onchocerciasis and provide them with a tool to help educate their clients as to their dogs' risk of developing this disease.

Comparison of bacterial culture results collected via direct corneal ulcer vs conjunctival fornix sampling in canine eyes with presumed bacterial ulcerative keratitis.

OBJECTIVE: To compare aerobic bacterial culture results between samples obtained from the corneal ulcer versus lower conjunctival fornix in eyes with presumed bacterial ulcerative keratitis. ANIMALS STUDIED: Fifty five client-owned dogs diagnosed with ulcerative keratitis. PROCEDURES: Ophthalmic examinations were performed on each dog including slit-lamp biomicroscopy and indirect ophthalmoscopy. Microbial swabs were collected by direct sampling of the infected corneal ulcer as well as the lower conjunctival fornix, of the same eye, using a sterile rayon-tipped swab. Samples were submitted to an outside reference laboratory for aerobic bacterial culture and sensitivity. RESULTS: One hundred twelve samples were obtained from 56 eyes (55 dogs). Sixty-eight samples yielded bacterial growth. Positive growth from both sites was obtained in 31 eyes (55%). Six eyes yielded bacterial growth from the conjunctival fornix but not from the cornea. No bacterial growth was obtained from either sampling site in 19 eyes. Overall, 31/56 (55%) corneal samples were positive and 37/56 (66%) conjunctival fornix samples were positive. Comparison of organisms isolated from the two collection sites of the same eye revealed an exact correlation in 42/56 (75%) eyes and differed in 14/56 (25%) eyes. Twenty different bacterial isolates were obtained from 68 positive samples. Gram-positive (71%) organisms were more common than Gram-negative (29%). The most commonly isolated organisms were Staphylococcus pseudintermedius (25%), beta-hemolytic Streptococcus spp. (23%), and Pseudomonas aeruginosa (12%). Methicillin-resistant organisms were isolated in 9% of samples. CONCLUSION: Sampling from the conjunctival fornix may be a suitable alternative to direct ulcer sampling in eyes with compromised corneal structural integrity.

Cofactors associated with Sudden Acquired Retinal Degeneration Syndrome: 151 dogs within a reference population.

OBJECTIVE: To determine factors associated with sudden acquired retinal degeneration syndrome (SARDS) diagnosed within one referral population. ANIMALS STUDIED: 151 dogs diagnosed with SARDS. PROCEDURES: Breed, age, sex, and body weight were compared between dogs with electroretinogram-confirmed SARDS and dogs presented to the UC Davis Veterinary Medical Teaching Hospital (UCD-VMTH) from 1991 to 2014. RESULTS: SARDS was diagnosed in 151 dogs, representing 1.3% of dogs presented to the UCD-VMTH for ophthalmic disease. Although dogs of 36 breeds were affected, the Dachshund (n = 31, 21%), Schnauzer (16, 11%), Pug (11, 7%), and Brittany (5, 3%) were significantly overrepresented, and the Labrador Retriever (3, 2%) was significantly underrepresented vs. the reference population (P < 0.001). Median (range) age and body weight of affected vs. reference dogs were 8.9 (3-20) vs. 6.8 (0.1-26) years and 12.4 (2.8-52.7) vs. 22.3 (0.1-60) kg, respectively. Dogs 6-10 years of age and between 10-20 kg in body weight were significantly overrepresented in the SARDS population, while dogs <6 years of age were significantly underrepresented (P < 0.01). Spayed females (59% of affected dogs) were significantly overrepresented compared to the reference population, whereas intact females (1% of affected dogs) were significantly underrepresented. CONCLUSIONS: Consistent with previous studies, smaller, middle-aged, spayed female dogs may be at increased risk of developing SARDS. Unlike previous studies, this is the first study comparing a variety of SARDS-affected breeds to a reference population. Potentially increased risk of SARDS in several breeds, particularly Dachshunds, suggests a familial factor that warrants further investigation using genetic techniques.

LupiQuant: A real-time PCR based assay for determining host-to-parasite DNA ratios of Onchocerca lupi and host Canis lupus from onchocercosis samples.

Onchocerca lupi is a filarial nematode that causes ocular onchocercosis in canines globally including North America and areas of Europe, North Africa, and the Middle East. Reported incidence of this parasite in canines has continued to steadily escalate since the early 21st century and was more recently documented in humans. Whole genome sequencing (WGS) of this parasite can provide insight into gene content, provide novel surveillance targets, and elucidate the origin and range expansion. However, past attempts of whole genome sequencing of other Onchocerca species reported a substantial portion of their data unusable due to the variable over-abundance of host DNA in samples. Here, we have developed a method to determine the host-to-parasite DNA ratio using a quantitative PCR (qPCR) approach that relies on two standard plasmids each of which contains a single copy gene specific to the parasite genus Onchocerca (major body wall myosin gene, myosin) or a single copy gene specific to the canine host (polycystin-1 precursor, pkd1). These plasmid standards were used to determine the copy number of the myosin and pkd1 genes within a sample to calculate the ratio of parasite and host DNA. Furthermore, whole genome sequence (WGS) data for three O. lupi isolates were consistent with our host-to-parasite DNA ratio results. Our study demonstrates, despite unified DNA extraction methods, variable quantities of host DNA within any one sample which will likely affect downstream WGS applications. Our quantification assay of host-to-parasite genome copy number provides a robust and accurate method of assessing canine host DNA load in an O. lupi specimen that will allow informed sample selection for WGS. This study has also provided the first whole genome draft sequence for this species. This approach is also useful for future focused WGS studies of other parasites.