Shiv Kumar Verma

New Jersey Department Of Health · Public Health and Environmental Laboratories

To assess the role of white-tailed deer (Odocoileus virginianus, WTD) in the epidemiology of toxoplasmosis, we conducted a national survey of WTD across the USA for Toxoplasma gondii infection. To do this, we combined serology with parasite isolation to evaluate the prevalence and genetic diversity of T. gondii in this game species. From October 20...

Genotyping of viable Toxoplasma gondii from the first USA national survey of feral swine revealed evidence for sylvatic transmission cycle, and presence of highly virulent parasite genotypes - J. P. Dubey, C. K. Cerqueira-Cézar, F. H. A. Murata, S. K. Verma, O. C.H. Kwok, K. Pedersen, B. M. Rosenthal, C. Su

In a national survey of fresh, unfrozen, American pasture-raised lamb and pork, the prevalence of viable Toxoplasma gondii was determined in 1500 samples selected by random multistage sampling (750 pork, 750 lamb) obtained from 250 retail meat stores from 10 major geographic areas in the USA. Each sample consisted of a minimum of 500 g of meat purc...

The protozoan parasite Sarcocystis falcatula is an important cause of clinical disease in several avian intermediate hosts. The host range of S. falcatula is wide and numerous outbreaks of acute sarcocystosis have been reported in passerine and psittacine birds in captivity in the Americas. Previous diagnosis was performed by serologic methods, lig...

Most eukaryotic parasites are obligately heteroxenous, requiring sequential infection of different host species in order to survive. Toxoplasma gondii is a rare exception to this rule, having a uniquely facultative heteroxenous life cycle. To understand the origins of this phenomenon, we compared development and stress responses in T. gondii to tho...

pH vs Ctl RNAseq data. Xls file containg raw reads, rLog values, and results for DESeq two analysis for pH vs control treated H. hammondi Amer and T. gondii VEG HFF infection.

Primer Table. File containing all of the primers used in this study, the purpose of the primer, and the Toxodb.org gene ID for each gene queried.

D4D15 RNASeq and 24 hr THP-1 RNAseq. Xls file containing raw reads, rLog values, and results for DESeq2 analysis for D4 and D15 HFF infection with H. hammondi Eth1 and T.gondii VEG and 24 hr THP-1 infection with H. hammondi Amer and T. gondii VEG.

Gene Set Categorization Significant Genes D4 vs D15 and 24 hr THP-1. Xls file containing all genes belonging to gene sets used in analyses and indicating which genes are significant in which categories for analysis. Significant genes for D4, D15, and 24 hr THP-1 data sets are indicated by the number 1, highlighted in a green box.

Gene Set Categorization Significant Genes pH vs Ctl. Xls file containing all genes belonging to gene sets used in analyses and indicating which genes are significant in which categories for analysis. Significant genes for pH versus control data sets are indicated by the number 1, highlighted in a green box.

Toxoplasmosis in marine mammals is epidemiologically and clinically important. Toxoplasma gondii antibodies (by modified agglutination test, cut-off ≥1:25) were detected in serum of 65 of 70 (92.9%) northern sea otters (Enhydra lutris kenyoni) from Washington State, USA. Brains and/or muscles of 44 sea otters were bioassayed in mice (INF-γ knock-ou...

Rodents are intermediate hosts for many species of Sarcocystis. Little is known of Sarcocystis cymruensis that uses the Brown rat (Rattus norvegicus) as intermediate hosts and the domestic cat (Felis catus) as experimental definitive host. Here, we identified and described Sarcocystis cymruensis in naturally infected R. norvegicus from Grenada, Wes...

Bovine besnoitiosis, caused by Besnoitia besnoiti, is an economically important disease of cattle in many countries but its transmission remains a mystery. Wild felids are suspected to be its definitive hosts. The domestic cat (Felis catus) is known experimental definitive host for Besnoitia species of rodents. Here, we report for Besnoitia darling...

The protozoan parasite Sarcocystis neurona is an important cause of disease in horses (equine protozoal myeloencephalitis, EPM) and marine mammals. Isolated reports of clinical EPM-like disease have been documented in a zebra, raccoon, domestic cat, domestic dog, ferret, skunk, mink, lynx, red panda and fisher. The predominant disease is encephalom...

Here we report a new species of Sarcocystis with Barred owls as the natural definitive host and interferon gamma gene knockout (KO) mice as an experimental intermediate host. A Barred owl submitted to the Carolina Raptor Center, Huntersville, North Carolina, was euthanized because of paralysis. Fully sporulated 12.5 x 9.9 m sporocysts were found i...

In the search for potential vaccine candidates for the control of human lymphatic filariasis, we recently identified calponin-like protein, that regulates actin/myosin interactions, in a proinflammatory fraction F8 (45.24–48.64 kDa) of Brugia malayi adult worms. In the present study, the gene was cloned, expressed, and the recombinant Calponin of B...

The muscles of herbivores commonly harbor sarcocysts of parasites belonging to species in the genus Sarcocystis, but such muscle parasites are rare in carnivores. Here, we report Sarcocystis arctica-like sarcocysts in muscles of Arctic foxes (Vulpes lagopus) from Alaska, USA, for the first time. The tongues of 56 foxes were examined for Sarcocystis...

Here, we report a new species of Sarcocystis with red-tailed hawk (RTH) as the natural definitive host and IFN-γ gene knockout (KO) mice as an experimental intermediate host in which sarcocysts form in muscle. Two RTHs submitted to the Carolina Raptor Center, Huntersville, North Carolina, were euthanized because they could not be rehabilitated and...

Here we report a new species, Sarcocystis pantherophisi n. sp. with the Eastern rat snake (Pantherophis alleghaniensis) as natural definitive host and the interferon gamma gene knockout (KO) mouse as the experimental intermediate host. Sporocysts (n = 15) from intestinal contents of the snake were 17.3 x 10.9 µm. Sporocysts were orally infective to...

Hammondia hammondi is the nearest relative of Toxoplasma gondii, but unlike T. gondii is obligately heteroxenous. We have compared H. hammondi and T. gondii development in vitro and identified multiple H. hammondi -specific growth states. Despite replicating slower than T. gondii , H. hammondi was resistant to pH-induced tissue cyst formation early...

Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. T. gondii has been reported in multiple cervid species; however, little is known of the parasite in North American elk (Cervus...

More than 200 valid Sarcocystis species have been described in the parasitological literature. The developmental life cycle in the intermediate host and definitive host has only been described for a few species. Sarcocystis parasites are common pathogens infecting a wide range of animals, including humans, and this unit reviews the methods used for...

Raptors serve as the definitive host for several Sarcocystis species. The complete life cycles of only a few of these Sarcocystis species that use birds of prey as definitive hosts have been described. In the present study, Sarcocystis species sporocysts were obtained from the intestine of a Cooper's hawk (Accipiter cooperii) and were used to infec...

Chickens are considered important in the epidemiology of Toxoplasma gondii. Chicken hearts (n = 1185) obtained from grocery stores were tested for T. gondii infection. Antibodies to T. gondii were assayed in fluid removed from the heart cavity using the modified agglutination test (MAT) at 1:5, 1:25, and 1:100 dilutions. MAT antibodies were detecte...

Author summary Toxoplasma gondii is an intracellular parasite that infects up to one-quarter of humans worldwide. Although it can infect virtually any warm-blooded animal, its definitive host is the cat where the sexual cycle occurs in enterocytes of the small intestine, producing microscopic, durable oocysts that are shed in feces and can remain i...

Strains and clones used in this study. (PDF)

Plasmids used in this study. (PDF)

Primers and plasmids used in this study. (PDF)

Number of enteroepithelial stages in HE-stained histological sections of cat ileum. (PDF)

Toxoplasma gondii causes lifelong chronic infection in both feline definitive hosts and intermediate hosts. Multiple exposures to the parasite are likely to occur in nature due to high environmental contamination. Here, we present data of high seroprevalence and multiple T. gondii strain co-infections in individual bobcats (Lynx rufus). Unfrozen sa...

There is considerable confusion concerning the identity of macroscopic Sarcocystis species in camels. Currently 2 species: Sarcocystis cameli, and Sarcocystis ippeni are recognized from 1-humped camel (Camelus dromedarius), and sarcocysts of both species are microscopic. Here, we report identity of macroscopic sarcocysts from the C. dromedarius in...

The objectives of the present cross sectional study were to isolate and genotype Toxoplasma gondii in free-range chickens from Grenada, West Indies. Using the modified agglutination test, antibodies to T. gondii were found in 39 (26.9%) of 145 free-range chickens with titers of 25 in 7chickens; 50 in 6 chickens; 100 in 2 chickens; 200 or higher in...

Sarcocystis neurona is the most frequent cause of equine protozoal myeloencephalitis, a debilitating neurological disease of horses that can be difficult to treat. We identified SnCDPK1, the S. neurona homologue of calcium-dependent protein kinase 1 (CDPK1), a validated drug target in Toxoplasma gondii. SnCDPK1 shares the glycine “gatekeeper” resid...

We created and tested multi-epitope DNA or protein vaccines with TLR4 ligand emulsion adjuvant (gluco glucopyranosyl lipid adjuvant in a stable emulsion [GLA-SE]) for their ability to protect against Toxoplasma gondii in HLA transgenic mice. Our constructs each included 5 of our best down-selected CD8+ T cell-eliciting epitopes, a universal CD4+ he...

There is an emerging concern that snakes are definitive hosts of certain species of Sarcocystis that cause muscular sarcocystosis in human and non-human primates. Other species of Sarcocystis are known to cycle among snakes and rodents, but have been poorly characterized in the USA and elsewhere. Although neurological sequalae are known for certain...

Raptors are good indicators of the prevalence of Toxoplasma gondii in the environment because they prey on small mammals and birds. These prey species are a major source of infection in domestic cats ( Felis catus ), which shed the environmentally resistant oocysts. We assessed T. gondii infection in 281 opportunistically available raptors at a reh...

Background Lymphatic filariasis caused by Wuchereria bancrofti, Brugia malayi and B. timori, is a debilitating disease with an adverse social and economic impact. The infection remains unabated in spite of treatment with existing antifilarial drugs diethylcarbamazine (DEC) and ivermectin which are chiefly microfilaricides. There is therefore, need...

Toxoplasma gondii, the most common parasitic infection of human brain and eye, persists across lifetimes, can progressively damage sight, and is currently incurable. New, curative medicines are needed urgently. Herein, we develop novel models to facilitate drug development: EGS strain T. gondii forms cysts in vitro that induce oocysts in cats, the...

Background: The protozoan parasite Toxoplasma gondii is one of the most widely distributed and successful parasites. Toxoplasma gondii alters rodent behavior such that infected rodents reverse their fear of cat odor, and indeed are attracted rather than repelled by feline urine. The location of the parasite encysted in the brain may influence this...

Wild birds are important in the epidemiology of toxoplasmosis because they can serve as reservoir hosts, and vectors of zoonotic pathogens including Toxoplasma gondii. Canada goose (Branta canadensis) is the most widespread geese in North America. Little is known concerning T. gondii infection in both migratory, and local resident populations of Ca...

Sarcosystis spp. are protozoan parasites found all over the world infecting a wide range of wild and domesticated animals including humans. Livestock diseases from these parasites cause the condemnation or downgrading of meat, poor growth, reduced milk, reduced wool, and animal death, resulting in millions of dollars of loses annually. Additionally...

Sarcocystis sarcocysts are common in muscles of herbivores but are rare in muscles of carnivores. Here, we report sarcocysts in the muscles of a gray wolf (Canis lupus) from Alaska, USA, for the first time. Sarcocysts extracted from the tongue of the wolf were up to 900 μm long and slender and appeared to have a relatively thin wall by light micros...

Paraffin-embedded blocks of brain of a nine months old bull calf that died of neurological signs in 1982 in Germany were restudied. Numerous schizonts and merozoites were found associated with extensive but focal necrosis and severe meningoencephalitis. Developing stages of schizonts as well as free merozoites were identified. The schizonts were pr...

Little is currently known of clinical toxoplasmosis in humans and animals in the Caribbean. We investigated the prevalence of IgG and IgM antibodies in 437 pregnant women from 10 English speaking Caribbean countries. Overall, antibodies (IgG) to Toxoplasma gondii (modified agglutination test, MAT, cut-off 1:6) were found in 174 (39.8 %) of 437 huma...

Toxoplasma gondii is a protozoan parasite transmitted by domestic cats ( Felis catus ) that has historically caused mortality in native Hawaiian birds. To estimate how widespread exposure to the parasite is in nene (Hawaiian Geese, Branta sandvicensis), we did a serologic survey for T. gondii antibody and genetically characterized parasite DNA from...

There is considerable confusion concerning the species of Sarcocystis in South American camelids (SAC). Several species names have been used; however, proper descriptions are lacking. In the present paper, we redescribe the macroscopic sarcocyst forming Sarcocystis aucheniae and describe and propose a new name, Sarcocystis masoni for the microscopi...

Toxoplasma gondii infections are widespread in white-tailed deer (Odocoileus virginianus) but little is known of its prevalence in other cervids in the USA. Moose (Alces alces) is a popular large game animal, hunted for its meat and trophy antlers. Here, we report seroprevalence, isolation, and genetic characterization of T. gondii from moose from...

Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut-off 1:25). Twenty nine of 50 bobcats and 1...

Wild felids are thought to share parasites with domestic cats. However, little is known of the coccidian parasites of wild felids. We investigated the presence of Sarcocystis spp. in tissues of 6 species of 90 Neotropical small felids killed in road accidents in the state of Rio Grande do Sul, Brazil by using microscopic and molecular techniques. F...

Four Roller pigeons (Columba livia f. dom.) at the Philadelphia Zoo died suddenly. Necropsy examination revealed macroscopic hepatitis. Microscopically, the predominant lesions were in liver, characterized with necrosis and mixed cell inflammatory response. Sarcocystis calchasi-like schizonts and free merozoites were identified in liver. Transmissi...

There is considerable confusion concerning Sarcocystis species in equids. Little is known of Sarcocystis infections in donkeys (Equus asinus). Here we describe the structure of Sarcocystis bertrami-like from the donkey by light microscopy (LM) and transmission electron microscopy (TEM). Nineteen sarcocysts from the tongue of a donkey from Egypt wer...

Infection with Sarcocystis is common in many species of wild cervids but none is reported from the black-tailed deer (Odocoileus hemionus columbianus). Here, we report Sarcocystis infection in two black-tailed deer from northwest USA for the first time. Sarcocysts were microscopic, up to 556 μm long and mature. The sarcocyst wall was up to 1.39 μm...

The North American opossum (Didelphis virginiana) is the definitive host for at least three named species of Sarcocystis: Sarcocystis falcatula, Sarcocystis neurona and Sarcocystis speeri. The South American opossums (Didelphis albiventris, Didelphis marsupialis and Didelphis aurita) are definitive hosts for S. falcatula and S. lindsayi. The sporoc...

Numerous species of Sarcocystis have been reported from wild ruminants, but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of three of seven mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; one had Sarcocystis cornagliai-like sarcocysts,...

Cattle (Bos taurus) are intermediate hosts for four species of Sarcocystis, namely Sarcocystis cruzi, Sarcocystis hirsuta, Sarcocystis hominis, and Sarcocystis rommeli. Of these four species, mature sarcocysts of S. cruzi are thin-walled (<1 μm), whereas S. hirsuta, S. hominis, and S. rommeli have thick walls (4 μm or more). Here, we describe a new...

There is considerable confusion concerning Sarcocystis species in camels. Five species: Sarcocystis cameli, Sarcocystis ippeni, Sarcocystis camelicanis, Sarcocystis camelocanis and Sarcocystis miescheri were named with inadequate descriptions and no type specimens. Here, we review literature on sarcocystosis in camels worldwide and redescribe struc...

Cattle (Bos taurus) are intermediate hosts for three named species of Sarcocystis, S. cruzi, S. hirsuta, and S. hominis. Recently, a fourth species was identified and named S. sinensis. However, S. sinensis was originally named for a species of Sarcocystis in water buffalo (Bubalus bubalis) in China. Based on unverifiable evidence, it was suggested...

This study compared genetic diversity of Toxoplasma gondii isolates from Portugal, Austria and Israel. For this, we genotyped 90 T. gondii isolates (16 from Portugal, 67 from Austria and 7 from Israel) using 10 nested PCR-restriction length polymorphism (RFLP) genetic markers and 15 microsatellite (MS) markers. By PCR-RFLP typing, 7 isolates from P...

Toxoplasma gondii infects virtually all warm-blooded hosts worldwide. Recently, attention has been focused on the genetic diversity of the parasite to explain its pathogenicity in different hosts. It has been hypothesized that interaction between feral and domestic cycles of T. gondii may increase unusual genotypes in domestic cats and facilitate t...

SUMMARY Transmission of pathogens between domestic and wild life animals plays an important role in epidemiology. Feral pig populations are increasing and expanding in the USA, and may constitute a risk to non-biosecure domestic pig facilities by serving as reservoirs for pathogens. We surveyed, for Sarcocystis infection, the myocardium of 1006 fer...

The consumption of unpasteurized goat cheese and goat's milk has been suggested as a risk factor for toxoplasmosis in humans. In the present study, detection and survival of Toxoplasma gondii in milk and cheese was studied by bioassay in mice (milk) and in cats (cheese). Eight goats were inoculated orally with 300 to 10,000 oocysts of T. gondii str...

There are several reports of Sarcocystis sarcocysts in muscles of dogs but these species have not been named. Additionally, there are 2 reports of Sarcocystis neurona in dogs. Here, we propose 2 new names, Sarcocystis caninum, and Sarcocystis svanai for sarcocysts associated with clinical muscular sarcocystosis in 4 domestic dogs (Canis familiaris)...

Previous studies have stressed the genetic divergence and high pathogenicity of strains of T. gondii from French Guiana. Although strains from coastal, human adapted environments (so called anthropized) resemble those found in other regions of the Caribbean, strains collected from inland jungle environment are genetically quite diverse. To better u...

Four valid species of Sarcocystis have been reported from the water buffalo (Bubalus bubalis): Sarcocystis fusiformis, Sarcocystis buffalonis, Sarcocystis levinei and Sarcocystis dubeyi. Here, we redescribe structure of S. fusiformis sarcocysts by scanning and transmission electron microscopy (SEM, TEM). Twenty-one macroscopic sarcocysts from oesop...