View
224
Download
0
Category
Preview:
Citation preview
Onchocerca lupi
Canid Falariasis in NM
BY AUDREY MELTON
“Coco”, inside/outside dogJemez mountains NM
Red inflamed eyesVeterinary treatmentBlindnessEuthanasia
AN EMERGING INFECTIOUS DISEASE IN DOGSNEW MEXICO
Veterinarians encounter“dog eye worms”
Bilateral Conjunctival swellingRetinal detachment
Surgical removal long thread-like worms
Filaria, NematodaOnchocerca sp.
Commonly O. lupi is mistaken as Thelazia callipaeda. Which is another “eyeworm” found in Canines in NM. They can look similar and cause a similar cyst/mass in the eye.
Adult worms can appear similar as well as the mass/cyst that forms in the eye
Life cycle of Thelazia callipaeda
Deposited in parasite collection ofMuseum for Southwestern Biology
(MSB)Perpendicular cuticular ridges(EtOH fixed specimen)
Morphology, ND5, CO1, obligate Wolbachia endosymbiont, Europe, (North America)
>lcl|OlupiNMCO1 Onchocerca lupiTTGGATGTTGCCTTTGATGTTGGGGGCTCCAGAGATGGCTTTTCCTCGAATTAACGCTTTGTCTTTTTGGTTTACTTTTGTATCTTTGTTGATGGTTTATCAGTCTTTTTTTATTGGAGGCGGTCCTGGTAGTAGTTGAACTTTTTATCCTCCTCTTAGAGTAGAGGGTCAGCCTGAGTTATCTTTGGATACTATAGTTTTGGGTTTGCATACTGTTGGTGTTGGTTCTTTGTTGGGTGCTATTAATTTTATGGTTACTACTCAAAATATGCGTTCTACTGCTGTGACTTTAGATCAAATTAGTATGTTTGTCTGGACTTCTTATTTAACTTCTTTTTTATTAGTATTATCTGTTCCTGTTTTAGCTGGGTCTTTGTTATTTTTGTTGTTGGATCGTAATTTTAGTACTTCTTTTTATGATACTAAAAAAGGTGGTAATCCTTTGTTGTATCAGCACTTGTTTTGGTTTTTCGGTCATCCCGAGGTTTATGTTATTATTTTACCGGTTTTTGGTATTATTAGGGAGGCAGTTTTATTTTTGACTGATAAGGATCGTTTGTTTGGTCAAACTAGGATGACTTTTGCTTCTATTTGGATTGCTGTTTTGGGGACTTCTGTTTGGGGTCACCATATGTATACGGCTGGTTTGGA
>lcl|OlupiND5 Onchocerca lupiCTGCTCCTACTCCTGTTAGTTGTTTGGTTCATAGTAGGACTTTAGTTACCGCGGGAGTTATGTTAATAGATTGTTATGTTTATGTTTCTTTGAATTCTGATGTTTTGTCTTTTGTTTTTTATGTTGGTTTTTTTACTATGGTTTTTTCTGGTTTTTGTTCTTTAGTTGAGCAGGATGCTAAAAAGATTGTTGCTTTGAGTACTATGTCTCAGATTGGTTTTTGTTTTTTGGCTATTGGTAGGGGTTTACATTATTTATCTTATGTTCATATGATTGGGCATTCTTTTTTTAAGAGTTTGTTATTTATGCAAATGGGTTATTTAATTTTTATTAATTTTGGTCAGCAGGATTGTCGTGGTTATTCTTTCTTTGGTTTTTGTGCCCCTGTTTTGGTTCAGTTACAGATTTTTTTATGTGTTTTTAGTT
SEQUENCES FROM COCO WORM SAMPLES
>lcl|Wolb_Olupi_NM Wolbachia endosymbiont of Onchocerca lupi from New Mexico, USA, 16S ribosomal RNA gene, partial sequenceTAGTGGCAGACGGGTGAGTAATGTATAGGAATCTACCTAGTAGTACGGAACAATTGCTGGAAACGACAACTAATACCGTATACGCCCTACGGGGGAAAGATTTATTGCTATTAGATGAGCCTATATTAGATTAGCTAGTTGGTAGGGTAATGGCCTGCCAAGGCTATGATCTATAGCTGATCTGAGAGGATGATCAGCCACACTGGAACTGAGATACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGGAAGCTTGATCCAGCCATGCCGCGTGAGTGAAGAAGGCCTTTGGGTTGTAAAGCTCTTTCAGTGAGGAAGATAATGACGGTACCACAGAAGAAGTCCTGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGAGGGCTAGCGTTATTCGGAATTATTGGGCGTAAAGAGCACGTAGGCTGGTTAGTAAGTTAAAAGTGAAATTCCAAAGCTTAACTTTGGAATTGCTTTTAAAACTGCTGATCTAGAGGTTGAAAGAGGATAGAGGAATTCCTAGTGTAGAGGTGAAATTCGTAAATATTAGGAGGAACACCAGTGGCGAAGGCGTCTATCTGGTTCAAATCTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGAATGTTAAATATGGGGAGATTACTTTCTGTGTTACAGCTAACGCGTTAAACATTCCGCCTGGGGACTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAAAACCTTACCACTCCTTG
nematodes I 20 leavesOnchocerca from Portugal cytochrome oxidase subunit (COI) gene, partial cds, mitochondrial
Onchocerca from Turkey cytochrome oxidase subunit (COI) gene, partial cds, mitochondrialOnchocerca lupi isolate 05RD‐0652 cytochrome oxidase subunit (COI) gene, partial cds, mitochondrial
nematodes I 8 leaves
nematodes I 14 leaves
nematodes I 6 leaves
nematodes I 3 leaves
nematodes I 3 leaves
nematodes I 13 leaves
nematodes I 17 leaves
nematodes I 7 leaves
nematodes I 3 leaves
nematodes I2l leaves
Dipetolonema gracile mitochondrial partial COI gene for cytochrome oxidase subunits
COCO WORM CO1, ND5 GROUP WITH Onchocerca lupiAND YIELDS Wolbachia ENDOSYMBIONT 16S SEQUENCE
CO1,Distance-tree through BLAST,(100-98% identity)
PROPOSED LIFE CYCLE Onchocerca lupi
Microfilariae penetrate vector’s gut and migrate to thoracic muscles
Adults in subcutaneous nodulespresent in periocular tissues
Migrate to head and vector’s proboscis
Vector insect takes a bloodmeal L3 larvae enter bite wound
Subcutaneous tissues
L2 Larvae
L3 Larvae
Vector insect
(as yet unidentified)takes a bloodmealIngests microfilariae
Skin and subcutaneous tissues
Vector?/hosts
EARLY DIAGNOSIS OCULAR FILARIASIS IN DOGS
•Transmission dynamics, vector, habitat?• Prevention, Control?• How many dogs, prevalence?• Worm genetic diversity, origin?
•SKIN SNIP•EtOH FIXED•EXTRACT DNA•PCR DETECTION OFMICROFILARIA(DEAD, IVERMECTIN)•CytB (+) CONTROL
ND5 CO1
1 2 3 4 1 2 3 4Coco SKINSNIPS
Sample SKINSNIP to capture microfilaria
Chris Bauer, Kellie WillitsChris Bauer, Kellie WillitsNancy McLean
“Blanca”
CytB ND5
Worm fragment
PREVALENCE, INTERVIEW VETS, OWNERS, GENETIC DIVERSITYFOR CIRCUMSTANCES
HABITAT, VECTOR
CytB CytBND5 ND5
POUND Skinsnips
TRACKING FILARIASIS IN DOGS, NM
Onchocerca lupi, insights/conclusions
Morphology, DNA, Morphology, DNA, WolbachiaWolbachia identify identify Onchocerca lupiOnchocerca lupi
DNA extraction EtOH skinsnips challenging (inhibitors?)DNA extraction EtOH skinsnips challenging (inhibitors?) PCR protocol labor intensive, considering qPCR protocol labor intensive, considering q--PCRPCR
Transmission dynamics, vector, habitat?Transmission dynamics, vector, habitat?Also in arid conditions (drought) , less likely blackflies, mosqAlso in arid conditions (drought) , less likely blackflies, mosquitoes, perhaps ticks or biting uitoes, perhaps ticks or biting midges.midges.
Prevention, Control?Prevention, Control?Ivermectin treatment (heartworm) seems to kill microfilariaIvermectin treatment (heartworm) seems to kill microfilariaNovel detection of (obligate endosymbiont) Novel detection of (obligate endosymbiont) WolbachiaWolbachia USA) use antibiotics against adult USA) use antibiotics against adult wormsworms
How many dogs, prevalence? How many dogs, prevalence? >6 specific dogs, Aztec skin snips 3/53 (5%) since Fall 2012>6 specific dogs, Aztec skin snips 3/53 (5%) since Fall 2012
Worm genetic diversity? origin? high sequence similarity/identiWorm genetic diversity? origin? high sequence similarity/identity to other ty to other O. lupiO. lupi is USA, is USA, Europe.Europe.
Resources
Canid Ocular Filariasis in NM Davila KA, Jones C, McLean NJ, Adema CM*, Melton, AL
Recommended