Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay.

BACKGROUND: Toxocara cati, the cat roundworm, is a parasitic nematode that known to cause toxocariasis in intermediate hosts and humans. In this study, we characterized the dynamics of T. cati larvae migration in BALB/c mice after inoculation with eggs and ensured the migration detecting the larval DNA by a PCR. To evaluate the dynamics of larval migration and distribution, twenty-four BALB/c mice were orally inoculated with 2500 T. cati infective eggs and the visceral organs of the infected animals were examined by pepsin digestion and microscopic parasite counts, followed by PCR at day 1 to 28 post-inoculation. RESULTS: The PCR assays were successfully used for detection of T. cati larvae in tissue samples and T. cati larvae and the DNAs were found in the liver, lungs, heart, kidneys and the brain. We detected T. cati in 92.2% of tissue samples by PCR, 30% higher than the conventional pepsin digestion technique. CONCLUSION: Our findings demonstrated that the PCR assay is a sensitive and specific for the detection of T. cati larvae. Therefore, it could become a useful tool for the investigation of the dynamics of larval migration and Toxocara infection in murine model.

Is it canine DUSN?: Another view of retinopathies, some acquired, and others possibly "inherited": Another view of retinopathies, some acquired, and others possibly "inherited".

The term retinopathy has been used to group several heterogeneous retinal abnormalities that are clearly acquired or are suspected/proposed to be inherited. Some share characteristic focal/multifocal hyperreflective tapetal lesions with a dark center, and areas of non-tapetal depigmentation suggestive of patchy or diffuse outer retinal atrophy. Progression is variable, and some develop unilateral or bilateral fundus changes resembling the clearly inherited form of retinal degeneration referred to as PRA. In this Commentary and Review, we discuss the role of ocular larva migrans resulting in the entity we refer to as canine DUSN and suggest that it may be responsible for some of the retinal findings grouped under the retinopathy rubric that share this characteristic fundus lesion.

Larva Migrans of Baylisascaris potosis In Experimental Animals.

Occurrence of clinical signs by infection with Baylisascaris potosis, the roundworm of kinkajous ( Potos flavus), in mice, rats, and rabbit were studied, and the migration behavior of larvae in mice were compared with that of Baylisascaris transfuga, the roundworm of bears ( Ursus spp.). Three groups of 8 mice, 3 groups of 6 rats, and 3 groups of 2 rabbits were inoculated with either 10, 100, or 1,000 B. potosis eggs. The other 8 mice were inoculated with 1,000 B. transfuga eggs. Animals were monitored for the occurrence of clinical signs until 60 days postinoculation (DPI). The carcass, viscera, brain, and eyes of each of 6 mice inoculated with 1,000 eggs of B. potosis or B. transfuga at 60 DPI were removed individually, and the number of larvae was counted. One mouse inoculated with 100 B. potosis eggs showed rolling at 27 DPI, and 1 larva was found in the medulla oblongata of this mouse. No clinical signs were observed in the other mice or in the rats and rabbits. A mean of 387.2 larvae was recovered from mice inoculated with 1,000 B. potosis eggs, and a mean of 422.0 larvae from mice inoculated with 1,000 B. transfuga eggs. The highest number of larvae was recovered from the carcasses for both B. potosis and B. transfuga. In the viscera, higher numbers of B. transfuga larvae (mean 131.8) were seen than B. potosis larvae (mean 33.1). In the brain, only 1 larva was detected in 1/6 mice inoculated with 1,000 B. potosis eggs, whereas a mean of 21 larvae was detected in mice inoculated with 1,000 B. transfuga eggs. A few larvae (range 0-1) were detected in the eyes of both mice inoculated with B. potosis or B. transfuga eggs. The result indicated that B. potosis larvae do not show a higher tendency to migrate into the brain of mice than B. transfuga larvae. However, 1 mouse inoculated with 100 eggs had 1 larva in the central nervous system and showed a serious neurological sign. This result may underline a potential risk of B. potosis to cause neural larva migrans in humans.

A modified method for molecular identification of Baylisascaris transfuga in European brown bears (Ursus arctos).

Baylisascaris transfuga is a roundworm that has been reported worldwide in most bear species. In mammals and possibly humans, the larvae of B. transfuga can migrate in the tissues of aberrant hosts with larva migrans syndrome. The current study was performed to identify B. transfuga in faecal samples from free-ranging brown bears in the Tatra Mountains National Park in southern Poland. A commercial kit was used to extract genomic DNA directly from faecal samples. Additionally, a Chelex resin-based technique was successfully implemented to prepare a PCR template from eggs retrieved by flotation. Based on the flotation results of 32 collected faecal samples, the prevalence of B. transfuga was 15.6%. The parasite was confirmed in samples found to be positive during the initial flotation by a molecular assay using DNA isolated directly from faeces. The retrieved eggs were confirmed as B. transfuga after their DNA was extracted using the Chelex protocol. Based on PCR amplification and sequencing of a 413-bp segment of cytochrome c oxidase 1 (COI), the obtained sequence was 100% identical to the COI segment of B. transfuga after a BLAST comparison to the GenBank™ database. The current study includes the first molecular confirmation of B. transfuga in brown bears in the western part of the Carpathians. We show that direct extraction of parasite DNA from bear faeces is efficient for molecular assays. As an alternative, we present the effectiveness of a Chelex-based technique for fast and convenient DNA isolation from the difficult-to-disrupt eggs of B. transfuga for PCR. Molecular tests of parasite DNA extracted directly from faecal material have limits of detection related to the amount of eggs in the samples. Thus, using classical flotation to obtain eggs for PCR may increase the credibility of the results, particularly in cases with a low number of excreted eggs. The Chelex resin protocol has potential for application in studies of intestinal parasites in wildlife for which conventional flotation is routinely used for microscopy.

BAYLISASCARIS PROCYONIS LARVA MIGRANS IN TWO CAPTIVE NORTH AMERICAN BEAVERS (CASTOR CANADENSIS).

Baylisascaris procyonis larva migrans was diagnosed in two North American beavers ( Castor canadensis ) belonging to a zoological park in Clarke County, Georgia. Both beavers presented with neurological signs. One beaver died naturally and despite attempted treatment, the other beaver was euthanatized because of severe clinical signs and poor prognosis. Histologic evaluation of the beavers revealed evidence of parasitic migration characterized by several lesions, including eosinophilic granulomas in various organs, as well as necrotizing eosinophilic and lymphoplasmacytic to granulomatous polioencephalitis, leukoencephalitis and cervical leukomyelitis. This represents the first confirmed case of B. procyonis larva migrans in beaver and the first non-raccoon ( Procyon lotor ) host in the southeastern United States. This report highlights the need for clinicians and diagnosticians to consider baylisascariasis in animals with compatible clinical signs. Preventative measures should be considered for captive animals, because early diagnosis of B. procyonis is challenging, and treatment is often unrewarding.

Cerebral Larva Migrans Caused by Baylisascaris spp. in a Free-ranging North American Porcupine ( Erethizon dorsatum ).

A free-ranging North American porcupine ( Erethizon dorsatum ) from Utah, US, exhibited neurologic symptoms and was submitted for necropsy. Histologic examination of the brain revealed severe encephalitis with an intralesional nematode larva consistent with Baylisascaris spp. Neurologic larva migrans had not been reported in free-ranging porcupines, or from wildlife in Utah.

Prevalence of Baylisascaris Roundworm in Captive Kinkajous in Japan.

Baylisascaris potosis causes larva migrans in animals. The present study evaluated the prevalence of B. potosis in captive kinkajous ( Potos flavus ) and the ability of milbemycin to treat natural infections of B. potosis in 2 female wild-caught kinkajous. In 2012, fecal samples were collected from 16 kinkajous in 6 zoological gardens and 29 imported captive kinkajous from 4 pet traders in Japan. Although all samples from zoological gardens were negative, 8 kinkajous from traders were positive for Baylisascaris eggs, at least 4 of which were wild caught in the Republic of Guyana. No associated human illness was reported from any of the facilities. The 2 infected kinkajous received a single oral administration of Milbemycin® A Tablets, which delivers 0.69-0.89 mg/kg milbemycin oxime. Fecal examinations on days 14 and 30 were negative for Baylisascaris eggs. These results demonstrated that milbemycin oxime has possible anthelmintic efficacy against Baylisascaris roundworms in captive kinkajous. We conclude that Baylisascaris infections are highly prevalent in wild-caught kinkajous in Japan and that most of the infected kinkajous were imported from the Republic of Guyana.

Baylisascaris procyonis larva migrans in two white-headed lemurs (Eulemur albifrons) in Spain and response to treatment derived from a human pediatric protocol.

Baylisascaris procyonis is a well-known ascaridoid nematode that causes larva migrans in humans and many other animal species. The North American raccoon (Procyon lotor) is the definitive host, which has been successfully introduced in the past decades to other geographical regions around the world. Two white-headed lemurs (Eulemuralbifrons) from a Zoological Park in Lugo, Spain, developed severe neurological signs within a brief period after being transferred from exhibit and placed in close contact with three captive raccoons from the same zoo. One lemur was euthanized due to the severity of disease progression and histopathology revealed granulomatous inflammation and ascaridoid larvae in kidneys, lung, spleen and brain. Larvae were identified as B. procyonis larvae by real time PCR. In light of the results, the cage mate with similar neurological signs was put on an albendazole treatment regimen adapted from a human pediatric protocol. The aggressive anthelmintic treatment likely contributed to the arrest of clinical signs and recovery of some motor skills. Importantly, Baylisascaris procyonis infection might occur in wild raccoon populations in Spain.

Experimental infection of Mongolian gerbils with Baylisascaris potosis.

The present study evaluated the pathogenicity of Baylisascaris potosis, a newly described ascarid nematode, in Mongolian gerbils. Gerbils were infected with varying doses of either B. potosis or Baylisascaris transfuga embryonated eggs (100, 1,000, and 4,000) for 30 days postinfection (pi). Baylisascaris potosis-infected gerbils showed no clinical signs of disease; however, gerbils exposed to 1,000 and 4,000 B. transfuga eggs showed severe neurologic signs at 22-29 days and 14-15 days pi, respectively. Histopathologic examination revealed larvae and lesions in the intestine, lung, liver, and muscles of B. potosis-infected gerbils, but not in the brain, whereas B. transfuga larvae were found only in the brain and muscle. These results indicate that B. potosis larvae migrate through numerous organs and are associated with visceral larva migrans in gerbils, but less frequently migrate to the nervous system in gerbils than does B. transfuga .

Cerebral Baylisascaris larva migrans in a cynomolgus macaque (Macaca fascicularis).

An incidental, asymptomatic, focal inflammatory lesion was detected in brain cerebrum of an approximately 6-year-old, female cynomolgus macaque from a chronic toxicology study. No gross lesions were noted at necropsy. Microscopically, the lesion contained a cross-section of larvae approximately 70-80 µm in diameter, a centrally located intestine flanked on either side by large triangular excretory columns, and prominent single lateral cuticular alae. Mixed inflammatory cells of eosinophils, macrophages, and lymphocytes admixed with abundant connective tissue stroma and necrosis surrounded the larvae. Histochemical stains for trichrome revealed significant amount of fibrous connective tissue. The morphology of the larvae was compatible with Baylisascaris spp. Based on the microscopic and histochemical examination, a diagnosis of neural Baylisascaris spp. larva migrans was made.

Zoonotic diseases associated with free-roaming cats.

Free-roaming cat populations have been identified as a significant public health threat and are a source for several zoonotic diseases including rabies, toxoplasmosis, cutaneous larval migrans because of various nematode parasites, plague, tularemia and murine typhus. Several of these diseases are reported to cause mortality in humans and can cause other important health issues including abortion, blindness, pruritic skin rashes and other various symptoms. A recent case of rabies in a young girl from California that likely was transmitted by a free-roaming cat underscores that free-roaming cats can be a source of zoonotic diseases. Increased attention has been placed on trap-neuter-release (TNR) programmes as a viable tool to manage cat populations. However, some studies have shown that TNR leads to increased immigration of unneutered cats into neutered populations as well as increased kitten survival in neutered groups. These compensatory mechanisms in neutered groups leading to increased kitten survival and immigration would confound rabies vaccination campaigns and produce naïve populations of cats that can serve as source of zoonotic disease agents owing to lack of immunity. This manuscript is a review of the various diseases of free-roaming cats and the public health implications associated with the cat populations.

Visceral and neural larva migrans in rhesus macaques.

Large ascarid larvae within granulomas were noted histologically in the mesenteric and pancreatic lymph nodes of 13 of 21 rhesus macaques (Macaca mulatta) euthanized as part of an experimental viral pathogenesis study. In addition, 7 of the 13 monkeys had cerebral granulomas, which in 4 animals contained nematode larvae similar to those within the lymph nodes. Despite the lesions, the animals did not show clinical signs associated with the parasitic infections. Characteristics of the larvae included, on cross-section, a midbody diameter of approximately 60 to 80 mum, a centrally located and slightly compressed intestine flanked on either side by large triangular excretory columns, and prominent single lateral cuticular alae. The morphology of the larvae was compatible with Baylisascaris spp. Baylisascariasis is a well-described infection of animals and humans that is caused by migrating larvae of the raccoon roundworm, Baylisascaris procyonis. A similar species, B. columnaris, is found in skunks and can cause cerebrospinal nematodiasis, but most reported cases of baylisascariasis have been due to B. procyonis. Our macaques were born free-ranging on an island in the southeastern United States where raccoons, but not skunks, were found to be common inhabitants, indicating that B. procyonis was the most likely parasite involved. These cases are similar to the low-level or covert cases of Baylisascaris infection described to occur in humans and provide further evidence of the existence of this parasite in the southeastern United States.

Identification of gender-regulated genes in Ancylostoma braziliense by real-time RT-PCR.

Ancylostoma braziliense belongs to the family Ancylostomatidae and infects cats and dogs in various parts of the tropical world. It is also a zoonotic parasite causing cutaneous larva migrants in humans. There are very few, either biological or molecular, studies of this species. In this study, differential display was used to identify differentially expressed genes in male and female A. braziliense. Nineteen new sequences were identified and examined by real-time RT-PCR to confirm male-female specificity. Ten were more expressed in males, while two were more expressed in females. Molecules shown to be important in other host-parasite relationships were also found in this study.

Visceral and presumptive neural baylisascariasis in an orangutan (Pongo pygmaeus).

A 32.5-year-old female hybrid orangutan (Pongo pygmaeus) developed hind-limb stiffness that progressed to tetraparesis over 2 wk. Repeated diagnostic evaluations, including serial magnetic resonance imaging of the central nervous system, revealed nonspecific lesions involving both the deep white and gray matter with an intact blood-brain barrier. Multiple empirical treatments failed to produce improvement and the animal was humanely euthanized. Histology of a granuloma in the ileum contained a nematode parasite, most consistent with Baylisascaris procyonis. Additionally, neuropil vacuolization, rarefaction, astrocytic scarring, and an eosinophilic granuloma and lymphoeosinophilic perivascular cuffing in the brain were suggestive of nematode migration. These findings confirm the presence of visceral larval migrans and support the presence of neural larval migrans. This case report of Baylisascaris procyonis confirms the presentation for the first time in an ape and documents the difficulty in antemortem diagnosis of neural larval migrans.

Larva migrans by Baylisascaris transfuga: fatal neurological diseases in Mongolian jirds, but not in mice.

Raccoon roundworms (Baylisascaris procyonis) and other Baylisascaris species cause patent or latent larva migrans (LM) in a variety of mammals and birds, including humans. It is not clear whether LM by Baylisascaris transfuga, roundworms of bears, is associated with clinical neurological disorders. To clarify this issue, ICR and BALB/c mice as well as Mongolian jirds (Meriones unguiculatus) were orally inoculated with 2,000-5,000 embryonated eggs of B. transfuga. In mice, the ascarid caused symptomatic LM of limited extent and duration, whereas the infection was fatal in jirds; i.e., they exhibited general signs such as severe depression and emaciation on days 8-11 postinfection (PI) and died, or they developed progressive and fatal neurological disorders after day 14 PI. Histological examination showed B. transfuga larvae in the brain of all mice and jirds examined, and the larvae collected from them developed to a size comparable with that of B. procyonis. There existed, however, critical differences in host reactions against larvae localized in the brain of mice and jirds; B. transfuga larvae found in mice were surrounded by granulomatous reactions and immobilized, whereas larvae found in jirds were free from any host reaction and mobile, causing extensive malacia.

Neuropathological observation of rabbits (Oryctolagus cuniculus) affected with raccoon roundworm (Baylisascaris procyonis) larva migrans in Japan.

Larvae of the raccoon roundworm, Baylisascaris procyonis (B. procyonis) are a known cause of cerebrospinal larva migrans in animals and humans. The present paper described details of the central nervous lesion in the rabbits (Oryctolagus cuniculus) affected with B. procyonis larva migrans in Japan. Clinically affected animals showed neurological signs including circling, torticollis, tremor of head, or ataxic gait. The most characteristic pathological alterations were large malacic lesions associated with an activated astroglial proliferation which was seen at the corpus medullare in the cerebellum including the cerebellar peduncle. Moreover, focal malacic lesions with perivascular cuffing and infiltration by lymphocytes and heterophiles were scattered everywhere throughout the brain. In these lesions or normal-appearing areas away from obvious lesions, ascarid larvae, about a maximum 65-75 micro m in diameter, were recognized. Other prominent features were minute lesions (we call them migration tract-like lesions) composed of lymphocytes, hemosiderin-laden macrophages and reactive astrocytes scattering throughout the cerebrum. In this study, we demonstrated ascarid larvae in only eight out of 23 animals diagnosed as B. procyonis larva migrans. Since it is not always possible to detect the larvae, the possibility of B. procyonis larva migrans must be given serious consideration to the characteristic lesions described above.

Epidemiological aspects of the first outbreak of Baylisascaris procyonis larva migrans in rabbits in Japan.

Larva migrans caused by the common raccoon ascarid, Baylisascaris procyonis, is a zoonotic disease of critical importance in North America. Recently we encountered the first proven outbreak of this disease in Japan in domestic rabbits (Oryctolagus cuniculus) in a small wildlife park. In this park, raccoons (Procyon lotor) had been kept for 9 years, and one raccoon was donated to the park by a pet owner 8 weeks prior to the occurrence of an outbreak in rabbits. Of 12 total raccoons, three raccoons including the donated one shed B. procyonis eggs in the feces, and two of these positive raccoons were kept in metal mesh cages on wooden pedestals, 2 m distant from the rabbit enclosure. Circumstantial evidence indicates that the donated raccoon is the likely source of this outbreak. Treatment of the raccoons with an ascaricide and decontamination by extensive flaming of the cages and the contaminated dirt floor of the park achieved a transient disappearance of ascarids from all 12 enclosed raccoons. Three months after the control measures began, recurrent ascarid infection was detected in three young raccoons of less than 1.5 years of age. The potential risk of serious zoonosis by B. procyonis as well as the difficulty in a clearance of contaminated areas should be considered by pet owners and public health workers in Japan.

First outbreak of Baylisascaris procyonis larva migrans in rabbits in Japan.

A syndrome of progressive neurological signs was noticed in rabbits (Oryctolagus cuniculus) kept in a small wildlife park in mid-July 2000. Three out of 12 common raccoons (Procyon lotor) kept in this park were infected with Baylisascaris procyonis, and the larvae were found from affected rabbits. This outbreak is the first proven B. procyonis larva migrans in Japan, and the potential risk of serious zoonosis by this ascarid species should be considered by pet owners, veterinarians, physicians and public health authorities in this country as in North America where raccoons are endemic.