Metabolic Labeling of Dirofilaria immitis Third- and Fourth-Stage Larvae and Their Excretory-Secretory Products

1991 ◽  
Vol 77 (6) ◽  
pp. 950 ◽  
Author(s):  
Glenn R. Frank ◽  
Robert B. Grieve
2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Christopher C. Evans ◽  
Katherine M. Day ◽  
Yi Chu ◽  
Bridget Garner ◽  
Kaori Sakamoto ◽  
...  

Abstract Background The Mongolian jird (Meriones unguiculatus) has long been recognized as a permissive host for the filarial parasite Brugia malayi; however, it is nonpermissive to another filarial parasite, canine heartworm (Dirofilaria immitis). By elucidating differences in the early response to infection, we sought to identify mechanisms involved in the species-specific clearance of these parasites. We hypothesized that the early clearance of D. immitis in intraperitoneal infection of the jird is immune mediated and parasite species dependent. Methods Jird peritoneal exudate cells (PECs) were isolated and their attachment to parasite larvae assessed in vitro under various conditions: D. immitis and B. malayi cultured separately, co-culture of both parasites, incubation before addition of cells, culture of heat-killed parasites, and culture with PECs isolated from jirds with mature B. malayi infection. The cells attaching to larvae were identified by immunohistochemistry. Results In vitro cell attachment to live D. immitis was high (mean = 99.6%) while much lower for B. malayi (mean = 5.56%). This species-specific attachment was also observed when both filarial species were co-cultured, with no significant change from controls (U(9, 14) = 58.5, p = 0.999). When we replicated these experiments with PECs derived from jirds subcutaneously infected with B. malayi, the results were similar (99.4% and 4.72% of D. immitis and B. malayi, respectively, exhibited cell attachment). Heat-killing the parasites significantly reduced cell attachment to D. immitis (mean = 71.9%; U(11, 14) = 7.5, p < 0.001) while increasing attachment to B. malayi (mean = 16.7%; U(9, 15) = 20, p = 0.002). Cell attachment to both species was reduced when larvae were allowed a 24-h pre-incubation period prior to the addition of cells. The attaching cells were identified as macrophages by immunohistochemistry. Conclusions These results suggest a strongly species-dependent response from which B. malayi could not confer protection by proxy in co-culture. The changes in cell attachment following heat-killing and pre-incubation suggest a role for excretory/secretory products in host immune evasion and/or antigenicity. The nature of this attachment is the subject of ongoing study and may provide insight into filarial host specificity.


1987 ◽  
Vol 73 (2) ◽  
pp. 377 ◽  
Author(s):  
David Abraham ◽  
Meisen Mok ◽  
Marcia Mika-Grieve ◽  
Robert B. Grieve

1986 ◽  
Vol 60 (1) ◽  
pp. 61-64 ◽  
Author(s):  
M. Mok ◽  
D. Abraham ◽  
R. B. Grieve ◽  
C. B. Thomas

AbstractThird-stage and fourth-stageDirofilaria immitislarvae exhibited positive thermotaxis when placed in a thermal gradient. Negative thermotaxis was not observed. Positive thermotaxis may be important for the successful transmission and for directing third and fourth-stage larval migration toward predilection sites in the Host.


Parasitology ◽  
2006 ◽  
Vol 132 (4) ◽  
pp. 535-543 ◽  
Author(s):  
H. CRAIG ◽  
J. M. WASTLING ◽  
D. P. KNOX

The nature of the proteins which comprise thein vitroexcretory/secretory products (ES) of the fourth-stage larva (L4) and adultTeladorsagia circumcinctaare largely undefined, despite the fact that this nematode induces profound changes, in part related to parasite ES, in the cellular architecture of the glands lining the abomasal surface of infected sheep and goats. In this study, the protein components of L4 and adult ES were fractionated using 1D gel electrophoresis and the major protein bands, detected by Coomassie blue staining, excised from the gel and subjected to tryptic digest and subsequent mass spectrometric analysis. The resultant peptide mass fingerprints were used to identify 15 L4 and 13 adult ES proteins. Several proteins, such as globin and some metabolic enzymes, were present in both ES. L4 ES alone contained thioredoxin peroxidase, an enzyme that can detoxify free radicals resulting from host inflammatory responses to the parasite, a cysteine proteinase which may aid penetration of the gastric mucosa and 2 different galectins which may influence cell differentiation and morphogenesis. Adult ES contained a nucleoside diphosphate kinase homologue, an enzyme which has been linked to cellular changes and can affect liquid secretion and goblet cell degranulation.


2011 ◽  
Vol 47 (3) ◽  
pp. 161-169 ◽  
Author(s):  
Barton W. Rohrbach ◽  
Agricola Odoi ◽  
Sharon Patton

Surveillance data indicate that failures have been reported for virtually all heartworm prevention product categories. Resistance of third and fourth stage larvae of Dirofilaria immitis to macrocyclic lactones, lack of compliance, other unknown factors, or a combination of these reasons may be the cause of failure. A survey of members of a national hunting dog club was conducted to identify practices used to prevent canine heartworm infections. Questionnaires were completed by 708 dog owners. Year-round administration of heartworm preventive medication was reported by 208 (88%) respondents residing north of the 37th parallel. Dosing was based on the estimated weight of the dog by 54 (7%) respondents, 389 (55%) did not record the date prophylaxis was administered, and 89 (13%) observed the dogs spit out pills. Heartworm testing at least once per year was done by 556 (79%) respondents and test dates were spread throughout the year. Only 448 (64%) respondents tested newly acquired dogs for heartworm. These findings suggest that veterinarians should place a greater emphasis on the frequency and timing of heartworm diagnostic tests, the importance of weighing dogs, duration of administration, recording the date monthly heartworm prophylaxis is given, and observing dogs to ensure that oral medication is retained.


Acta Tropica ◽  
1990 ◽  
Vol 47 (5-6) ◽  
pp. 339-353 ◽  
Author(s):  
Alan L. Scott ◽  
M.Sofi Ibrahim ◽  
Wesley K. Tamashiro

1992 ◽  
Vol 75 (2) ◽  
pp. 213-222 ◽  
Author(s):  
Jennifer K. Richer ◽  
Judy A. Sakanari ◽  
Glenn R. Frank ◽  
Robert B. Grieve

1988 ◽  
Vol 65 (2) ◽  
pp. 157-167 ◽  
Author(s):  
David Abraham ◽  
Robert B. Grieve ◽  
Marcia Mika-Grieve

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 786
Author(s):  
Lucienne Tritten ◽  
Erica J. Burkman ◽  
Tobias Clark ◽  
Guilherme G. Verocai

The canine heartworm, Dirofilaria immitis, is among the most important parasites of dogs in the United States and worldwide, and may cause severe and potentially fatal disease. Current diagnostic recommendations rely on serological detection of an adult female antigen, and visualization of microfilariae in the blood. Therefore, a reliable diagnosis can be only performed approximately six months post-infection. There is a growing need to characterize novel diagnostic markers that are capable of detecting the early stages of heartworm infection, in special markers associated with third-stage larvae (L3) and fourth-stage larvae (L4). The early detection of infection would guide medical interventions that could impede the development of patent infections and further parasite transmission. We cultured D. immitis L3 and L4 of two laboratorial strains with different susceptibility statuses to macrocyclic lactone drugs in vitro. Excretory/secretory microRNAs were sequenced and analyzed. We identified two miRNA novel candidates secreted abundantly by both L3 and L4 of both strains. These candidates were previously detected in the secretions of other D. immitis stages and one of them was found in the blood of D. immitis-infected dogs. These miRNAs have not been found in the secretions of other nematodes and could be D. immitis-specific diagnostic biomarkers, which could allow for the early detection of infection.


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