A 10-year-old male spotted seal presented with loss of appetite and decreased activity. Grossly, the internal organs revealed several filarial nematodes in the right ventricle of the heart and the pulmonary vessels. Histopathological examination of the brain revealed moderate nonsuppurative meningoencephalitis with glial nodules and neuronophagia. Japanese encephalitis virus (JEV) of genotype I was isolated from the brain. All nematodes were identified as Dirofilaria immitis. This is the first clinical case of co-infection with D. immitis and JEV in a seal, suggesting that the seal, may be a dead-end host, like the human and horse, for JEV.
Appetite ; Asian Continental Ancestry Group ; Brain ; Child ; Coinfection ; Dirofilaria immitis ; Dirofilaria ; Encephalitis Virus, Japanese ; Encephalitis, Japanese ; Genotype ; Heart ; Heart Ventricles ; Horses ; Humans ; Male ; Meningoencephalitis ; Republic of Korea

An 8-year-old female Eurasian otter (Lutra lutra) reared in a wetland center, died 2 h after sudden onset of astasia and dyspnea despite medical treatment. Gross examination of internal organs revealed 10 adult filarioid nematodes in the right ventricle of the heart and three between the left and right cerebral hemispheres. All nematodes were identified as Dirofilaria immitis by direct microscopy and polymerase chain reaction assay. Histopathological observation revealed multifocal hemorrhage in the cerebral subarachnoid space and focal necrosis with hemorrhagein the cerebellar parenchyma. Although rare, veterinarians should consider cerebral dirofilariasis as a differential diagnosis in unexplained neurological cases.
Adult ; Animals ; Brain ; Cerebrum ; Child ; Diagnosis, Differential ; Dirofilaria immitis ; Dirofilaria ; Dirofilariasis ; Dyspnea ; Female ; Heart ; Heart Ventricles ; Hemorrhage ; Humans ; Korea ; Microscopy ; Necrosis ; Otters ; Polymerase Chain Reaction ; Subarachnoid Space ; Veterinarians ; Wetlands

The present study was performed to investigate the seroprevalence and risk factors for Dirofilaria immitis infection in cats from Liaoning province, northeastern China. From October 2014 to September 2016, sera of 651 cats, including 364 domestic cats and 287 feral cats (332 females and 319 males) were assessed. They were tested for the presence of D. immitis antigen using SNAP Heartworm RT test kit. In this population, the average prevalence was 4.5%. Age and rearing conditions (feral or domestic) were found to be associated with the prevalence of D. immitis. The prevalence was significantly higher in feral cats compared with domestic cats (8.4% vs 1.4%, P < 0.01). There was no significant difference between males and females (4.7% vs 4.2%, P>0.05), but older cats (≥3 years old) showed a statistically higher prevalence compared with younger cats ( < 3 years old) in feral populations (16.8 vs 2.4%, P < 0.01), while the difference between the age groups was not statistically significant in domestic cats (2.4% vs 0.51%, P>0.05), all these results suggest that outdoor exposure time may be one of the most important factors for D. immitis prevalence in cats. Results reveal that D. immitis are prevalence in domestic and feral cats in northeastern China, which indicates that appropriate preventive measures should be taken to decrease the incidence of feline heartworm disease in Liaoning province, northeastern China.
Animals ; Cats* ; China* ; Dirofilaria immitis* ; Dirofilaria* ; Dirofilariasis ; Female ; Humans ; Incidence ; Male ; Prevalence ; Risk Factors ; Seroepidemiologic Studies*

Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.
Aedes ; Anopheles ; Bacteria ; Culex ; Culicidae* ; Dirofilaria ; Dirofilaria immitis ; Dirofilaria repens ; Egypt* ; Hope ; Insects ; Larva* ; Multiplex Polymerase Chain Reaction ; Parasites ; Polymerase Chain Reaction ; Reproduction ; Wolbachia* ; Wuchereria bancrofti

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.
Aedes/*parasitology ; Animals ; Anopheles/*parasitology ; Culex/*parasitology ; Dirofilaria immitis/genetics/*isolation & purification ; Dirofilaria repens/genetics/*isolation & purification ; Egypt ; Entomology/methods ; Female ; Multiplex Polymerase Chain Reaction/*methods ; Parasitology/methods ; Sensitivity and Specificity ; Wuchereria bancrofti/genetics/*isolation & purification

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.
Aedes/*parasitology ; Animals ; Anopheles/*parasitology ; Culex/*parasitology ; Dirofilaria immitis/genetics/*isolation & purification ; Dirofilaria repens/genetics/*isolation & purification ; Egypt ; Entomology/methods ; Female ; Multiplex Polymerase Chain Reaction/*methods ; Parasitology/methods ; Sensitivity and Specificity ; Wuchereria bancrofti/genetics/*isolation & purification

This study was conducted to determine the serum cardiac troponin I (cTnI) concentrations in clinically normal (uninfected) Korean Jindo dogs and those infected with Dirofilaria (D.) immitis. Forty Korean Jindo dogs (22 females, 18 males) were obtained from two kennels in the Boryeong area of Chungnam Province and the Daejeon area, Korea. Significantly higher cTnI concentrations were observed in D. immitis-infected dogs than clinically normal dogs. cTnI testing in addition to thoracic radiography, echocardiography, and electrocardiography shows promise as an additional method for diagnosis of heartworm infection.
Animals ; Chungcheongnam-do ; Diagnosis ; Dirofilaria immitis* ; Dirofilaria* ; Dogs* ; Echocardiography ; Electrocardiography ; Female ; Humans ; Korea ; Radiography, Thoracic ; Troponin I* ; Troponin*

The purpose of this study was to conduct a survey of Dirofilaria immitis infection among stray cats in Korea using nested PCR. We included 235 stray cats (121 females and 114 males) and evaluated each for the presence of feline heartworm infection. Blood samples were collected from 135 cats in Daejeon, 50 cats in Seoul, and 50 cats from Gyeonggi-do (Province). Of the 235 DNA samples, 14 (6.0%) were positive for D. immitis. The prevalence of infection in male cats (8/114, 7.0%) tended to be higher than that in female cats (6/121, 5.0%), but the difference was not statistically significant. In each location, 8, 2, and 4 cats were positive for infection, respectively, based on DNA testing. No significant differences in the prevalence were observed among the geographic regions, although the rate of infection was higher in Gyeonggi-do (8.0%) than Daejeon (5.9%) and Seoul (4.0%). We submitted 7 of the 14 D. immitis DNA-positive samples for sequencing analysis. All samples corresponded to partial D. immitis cytochrome c oxidase subunit I gene sequences with 99% homology to the D. immitis sequence deposited in GenBank (accession no. FN391553). To the best of our knowledge, this is the first survey using nested PCR to analyze the prevalence of D. immitis in stray cats in Korea.
Animals ; Blood/parasitology ; Cat Diseases/*epidemiology/*parasitology ; Cats ; DNA, Helminth/chemistry/genetics ; Dirofilaria immitis/genetics/*isolation & purification ; Dirofilariasis/*epidemiology/*parasitology ; Electron Transport Complex IV/genetics ; Female ; Korea/epidemiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/methods ; Prevalence ; Sequence Analysis, DNA ; Sequence Homology

Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.
Animal Structures/chemistry ; Animals ; Antibodies, Helminth/blood ; Antigens, Helminth/chemistry/*genetics/immunology/*isolation & purification ; Cloning, Molecular ; Dirofilaria immitis/chemistry/*genetics/immunology ; Disease Models, Animal ; Dogs ; Escherichia coli/genetics ; Gene Expression ; Molecular Sequence Data ; Molecular Weight ; Recombinant Fusion Proteins/chemistry/genetics/immunology/isolation & purification ; Sequence Analysis, DNA ; Tumor Markers, Biological/chemistry/*genetics/immunology/*isolation & purification

No abstract available.
Aged ; Animals ; Diagnosis, Differential ; Dirofilaria immitis/*isolation & purification ; Dirofilariasis/*diagnosis/parasitology/surgery ; Eye Infections, Parasitic/*diagnosis/parasitology/surgery ; Female ; Humans ; Ophthalmologic Surgical Procedures ; Orbital Diseases/*diagnosis/parasitology/surgery