Antibodies to the ventral disc protein delta-giardin prevent in vitro binding of Giardia lamblia trophozoites.

A cDNA coding for detlaq-giardin was cloned from Giardia lamblia trophozoites to localize the protein and to study its function in mediating surface attachment. Recombinant delta-giardin antigen was expressed in Escherichia coli as a poly-histidine fusion protein and was purified by affinity chromatography for production of antisera to delta-giardin. By immunoblotting analysis, antisera to recombinant delta-giardin antigen recognized a 31-kDa protein on G.

A longitudinal study of Giardia duodenalis genotypes in dairy cows from birth to 2 years of age.

Fecal specimens were collected from 30 calves from birth to 24 months of age at a dairy farm in Maryland to determine the prevalence of Giardia duodenalis genotypes in cattle of different ages. Fecal samples were subjected to density gradient centrifugation to remove debris and concentrate cysts. Specimens were analyzed by immunofluorescence microscopy and polymerase chain reaction (PCR).

Molecular and immunohistochemical detection of assemblage E, Giardia duodenalis in scouring North Dakota calves.

Despite many reports on the shedding of Giardia parasites by scouring calves, the role of Giardia as a cause of calf diarrhea is still controversial. To elucidate the role of Giardia duodenalis in calf scours, diagnostic samples from 189 scouring calves were tested by different assays during a 1-year-study period. Giardia antigens were detected in 22/189 scouring calves by a fecal-based enzyme-linked immunosorbent assay and 10 of these were positive for assemblage E, G.

Cryptosporidium ryanae n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus).

A new species, Cryptosporidium ryanae, is described from cattle. Oocysts of C. ryanae, previously identified as the Cryptosporidium deer-like genotype and recorded as such in GenBank (AY587166, EU203216, DQ182597, AY741309, and DQ871345), are similar to those of Cryptosporidium parvum and Cryptosporidium bovis but smaller.

A longitudinal study of cryptosporidiosis in dairy cattle from birth to 2 years of age.

Fecal specimens were collected from 30 calves from birth to 24 months of age at a dairy farm in Maryland to determine the prevalence and age distribution of Cryptosporidium species/genotypes. After centrifugation to remove debris and concentrate oocysts, specimens were examined by immunofluorescence microscopy and polymerase chain reaction (PCR). Fragments of the SSU-rDNA gene amplified by PCR were purified and PCR products were sequenced.

Detection of Assemblage A, Giardia duodenalis and Eimeria spp. in alpacas on two Maryland farms.

Sixty-one fecal samples were collected from adult alpacas and crias (ages 10 weeks to 10 years) on two farms in central Maryland. The farms raised both suri (silky-haired) and huacaya (crimpy-haired) breeds. Females and crias were housed together on pasture, whereas older/breeding males were maintained on separate pastures.

Detection of Cryptosporidium parvum oocysts by dot-blotting using monoclonal antibodies to Cryptosporidium parvum virus 40-kDa capsid protein.

Monoclonal antibodies (MAb) were prepared against the 40-kDa capsid protein of Cryptosporidium parvum virus (CPV) by immunizing mice with purified recombinant CPV40 protein. In immunoblotting analysis, MAbCPV40-1 bound to a 40-kDa protein in extracts of C. parvum oocysts.

Prevalence of Giardia duodenalis genotypes in adult dairy cows.

The prevalence of Giardia duodenalis genotypes was determined in adult dairy cows. Fecal specimens were collected from two farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens, cleaned of fecal debris and concentrated using CsCl density gradient centrifugation, were subjected to PCR and DNA sequence analysis.

Prevalence and molecular characterization of Cryptosporidium and Giardia species and genotypes in sheep in Maryland.

In the United Kingdom and Australia sheep have been implicated as sources of Cryptosporidium and Giardia that infect humans, but no such studies have been conducted in North America. Therefore, a study was undertaken to investigate the prevalence of these parasites in sheep on a farm in Maryland. Feces were collected from 32 pregnant ewes 1, 2, and 3 days after parturition and from each of their lambs 7, 14, and 21 days after birth.

Prevalence of Microsporidia, Cryptosporidium spp., and Giardia spp. in beavers (Castor canadensis) in Massachusetts.

Feces from 62 beavers (Castor canadensis) in Massachusetts were examined by fluorescence microscopy (IFA) and polymerase chain reaction (PCR) for Microsporidia species, Cryptosporidium spp., and Giardia spp. between January 2002 and December 2004.

Giardia and Cryptosporidium species and genotypes in coyotes (Canis latrans).

Feces and duodenal scrapings were collected from 22 coyotes (Canis latrans) killed in managed hunts in northeastern Pennsylvania. Polymerase chain reaction (PCR) methods were used to detect Giardia and Cryptosporidium spp. PCR-amplified fragments of Giardia and Cryptosporidium spp.

Prevalence of Cryptosporidium species and genotypes in mature dairy cattle on farms in eastern United States compared with younger cattle from the same locations.

Feces collected from 541 milking cows on two dairy farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida were examined for the presence of Cryptosporidium oocysts. Oocysts were concentrated from 15 g of feces from each cow and DNA was extracted. A two-step nested PCR protocol was used to amplify an 830 base pair fragment of the SSUrRNA gene.

Cryptosporidium, Giardia and Enterocytozoon bieneusi in cats from Bogota (Colombia) and genotyping of isolates.

The prevalence of Cryptosporidium, Giardia, and Enterocytozoon bieneusi in cats from Bogota (Colombia) was determined from fecal specimens and scrapings of duodenal and ileal mucosa screened by PCR. All PCR-positive specimens were sequenced to determine the genotype(s) present. Of 46 cats, 6 (13%) were positive for Cryptosporidium, 5 (11%) were infected with C.

Prevalence and genotypes of Giardia duodenalis in 1-2 year old dairy cattle.

To determine the prevalence of Giardia genotypes in 12-24 month old dairy heifers, fecal specimens were collected from two farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens, cleaned of fecal debris and concentrated using CsCl density gradient centrifugation, were subjected to PCR and DNA sequence analysis. Prevalence of Giardia infection, ranged from 11% to 75% on 14 farms with an average prevalence of 36% (204 positive cattle out of 571 examined).

Detection of Cryptosporidium felis and Giardia duodenalis Assemblage F in a cat colony.

Eighteen cats, 3-6 months of age, bred and housed in a closed colony, were transferred from that colony and placed in separate stainless steel cages in a building designed for housing animals. At daily intervals, feces were collected from the litter pans in each cage, pans and cages were cleaned, and fresh food and water were provided. Beginning 4 weeks after the transfer, oocysts of Cryptosporidium were detected in the feces of two cats by brightfield microscopy.

Prevalence of species and genotypes of Cryptosporidium found in 1-2-year-old dairy cattle in the eastern United States.

The prevalence of Cryptosporidium species in 1-2-year-old heifers was determined for 571 animals on 14 dairy farms in seven states on the East Coast of the United States. A fecal specimen collected directly from each heifer was processed to concentrate oocysts that were then examined by polymerase chain reaction (PCR). For every PCR-positive specimen the 18S rRNA gene of Cryptosporidium was sequenced.

Prevalence and genotypes of Giardia duodenalis in post-weaned dairy calves.

To determine the prevalence of Giardia genotypes in post-weaned dairy calves, fecal specimens were collected from 3 to 11-month-old dairy calves per farm on two farms in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and DNA sequence analysis. Overall, PCR provided more sensitive detection than IFA.

Genotypes of Cryptosporidium species infecting fur-bearing mammals differ from those of species infecting humans.

Of 471 specimens examined from foxes, raccoons, muskrats, otters, and beavers living in wetlands adjacent to the Chesapeake Bay, 36 were positive for five types of Cryptosporidium, including the C. canis dog and fox genotypes, Cryptosporidium muskrat genotypes I and II, and Cryptosporidium skunk genotype. Thus, fur-bearing mammals in watersheds excreted host-adapted Cryptosporidium oocysts that are not known to be of significant public health importance.

Concentrating, purifying and detecting waterborne parasites.

There has been recent emphasis on developing better methods for detecting diseases of zoonotic and veterinary importance. This has been prompted by an increase in human disease agents detectable in environmental samples, the potential for bioterrorism, and the lowering of international trade barriers and expansion of personal travel, which are bringing previously considered exotic diseases to new geographical localities. To appreciate the complexities of developing detection methods and working with environmental samples, it is appropriate to review technologies currently in use, as well as those in development and presently limited to research laboratories.

Prevalence of Giardia duodenalis genotypes in pre-weaned dairy calves.

To determine the prevalence of Giardia genotypes in pre-weaned dairy calves, fecal samples were collected from a minimum of 18, 1-7-week-old dairy calves per farm on two farms each in the states of Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Samples cleaned of fecal debris and concentrated using CsCl density gradient centrifugation were stained and examined by immunofluorescence microscopy and also subjected to PCR and gene sequence analysis. Prevalence by PCR ranged from 9% on a farm in Pennsylvania to 93% on a farm in Vermont, with an average prevalence for 407 calves on 14 farms of 40%.

Prevalence and age-related variation of Cryptosporidium species and genotypes in dairy calves.

Fifteen dairy farms in seven states on the east coast of the US were each visited on two consecutive years to determinate the prevalence of Cryptosporidium species in pre-weaned (5 days to 2 months) and post-weaned calves (3-11 months), respectively. After each of 971 fecal specimens collected directly from each calf was sieved and subjected to density gradient centrifugation to remove debris and concentrate oocysts, specimens were examined by immunofluorescence microscopy, and polymerase chain reaction (PCR). For all PCR-positive specimens the 18S rRNA gene of Cryptosporidium was sequenced.

Prevalence of Enterocytozoon bieneusi in post-weaned dairy calves in the eastern United States.

Fecal specimens were obtained from 3- to 8-month-old post-weaned dairy calves on farms in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. After removal of fecal debris by sieving and density gradient centrifugation, 59 of 452 calves (13%) from 11 farms in six states were found positive for Enterocytozoon bieneusi by PCR and DNA sequence analysis. Based on gene sequence data this genotype of E.

Triosephosphate isomerase gene characterization and potential zoonotic transmission of Giardia duodenalis.

To address the source of infection in humans and public health importance of Giardia duodenalis parasites from animals, nucleotide sequences of the triosephosphate isomerase (TPI) gene were generated for 37 human isolates, 15 dog isolates, 8 muskrat isolates, 7 isolates each from cattle and beavers, and 1 isolate each from a rat and a rabbit. Distinct genotypes were found in humans, cattle, beavers, dogs, muskrats, and rats. TPI and small subunit ribosomal RNA (SSU rRNA) gene sequences of G.

Molecular characterization of microsporidia indicates that wild mammals Harbor host-adapted Enterocytozoon spp. as well as human-pathogenic Enterocytozoon bieneusi.

Over 13 months, 465 beavers, foxes, muskrats, otters, and raccoons were trapped in four counties in eastern Maryland and examined by molecular methods for microsporidia. A two-step nested PCR protocol was developed to amplify a 392-bp fragment of the internal transcribed spacer region of the rRNA gene of Enterocytozoon spp., with the use of primers complementary to the conserved regions of published nucleotide sequences.