Molecular identification of Giardia duodenalis isolates from humans, dogs, cats and cattle from the state of São Paulo, Brazil, by sequence analysis of fragments of glutamate dehydrogenase (gdh) coding gene

doi:10.1016/j.vetpar.2007.08.019

Veterinary Parasitology

Volume 149, Issues 3–4, 10 November 2007, Pages 258-264

https://doi.org/10.1016/j.vetpar.2007.08.019 Get rights and content

Abstract

The nucleotide sequence of glutamate dehydrogenase (gdh) coding genes were obtained from cysts of Giardia duodenalis isolated from feces of naturally infected cats (n = 19), dogs (n = 27), humans (n = 37) and cattle (n = 5). The samples were from several municipalities within the state of São Paulo, Brazil and were collected from January 2004 to August 2006. Sequences analysis of the 37 specimens recovered from humans revealed 29 G. duodenalis assemblage AII and 8 G. duodenalis assemblage B. Among samples from cats, 11 were classified into assemblage F and 8 into assemblage AI. Only the host-adapted assemblages C and D were detected in samples from dogs (7 and 20, respectively). Among the samples from cattle, the genotype livestock was found in four samples and the assemblage AI was detected in one sample. The molecular identification of assemblages of G. duodenalis isolates from different hosts reveals that genetic diversity of this protozoon in Brazil is similar to that of Giardias from other parts of the world.

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Acknowledgements

The authors extend thanks to FAPESP for financial support, grant number 03/01587-3, and to CNPq for a scholarship for productivity in research to Solange M. Gennari and Leonardo J. Richtzenhain.

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Cryptosporidium and Giardia in cats and dogs: What is the real zoonotic risk?
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Due to the close bond between humans and companion animals, a thorough understanding of the diversity of Cryptosporidium species and Giardia assemblages in cats and dogs is essential to determine the potential zoonotic risks. Analysis of molecular studies shows that C. felis and C. canis are the main species infecting cats and dogs, respectively. These species are largely host-specific, as despite intense association with humans, prevalence of C. felis and C. canis in humans is low and predominantly in immunocompromised individuals and low-income countries. There have been reports of C. parvum in cats and dogs and two reports of C. hominis in dogs. In most studies conducted to date, however, the prevalence of zoonotic species was low and may be associated with coprophagy and or/spillback, but this remains to be determined. Results of subtyping studies suggest that for C. felis and C. canis, some zoonotic transmission may occur but host-adapted subtypes also exist. Giardia duodenalis assemblages C and D are commonly reported in dogs, with assemblages F and A most common in cats. Assemblages C, D and F are largely host-specific as there are only a handful of reports of them in humans. Reports of assemblage A and B in cats and dogs may be due to coprophagy or spillback from owners. Despite the extent of pet ownership and the close contact between humans and companion animals worldwide, the overall risk of zoonotic transmission from cats and dogs to humans is uncertain but thought to be low due to C. canis, C. felis and G. duodenalis assemblages C, D and F being predominantly host-specific, the relatively low prevalence of C. parvum (and C. hominis) in cats and dogs (which may be due to mechanical carriage), and low oo/cyst shedding. Carefully designed epidemiological studies of cats and dogs and their owners using subtyping tools are essential to better quantify the extent of spillover and spillback of Cryptosporidium and Giardia between pets and their owners.
First report of Giardia duodenalis assemblage F in humans and dogs in southern Brazil
2022, Comparative Immunology, Microbiology and Infectious Diseases
The present study aimed to molecularly characterize Giardia duodenalis from stool samples of humans, dogs, and cats. Molecular analyses were performed on 59 samples that tested positive for G. duodenalis on coproparasitological examinations. After extraction, the samples were first tested by nested polymerase chain reaction (n-PCR) analysis of the SSU-rRNA gene, and for the samples that were positive, the β-giardin, TPI, and GDH genes were analyzed. The amplicons obtained in the n-PCR of the β-giardin gene were subjected to PCR-restriction length polymorphism (RFLP) analysis and subsequent digestion with the enzyme HaeIII to differentiate the assemblages. Seven (11.8 %), 34 (57.7 %), and 18 (30.5 %) out of 59 samples were from humans, dogs, and cats, respectively. Nested-PCR results showed that 49.2 % (29/59) of samples were positive for the SSU-rRNA gene, with 42.9 % (3/7) of humans, 55.9 % (19/34) of dogs, and 38.9 % (7/18) of catsve. Of the other genes analyzed, β-giardin was amplified most frequently, in 34.5 % (10/29) of samples, followed by GDH in 27.6 % (8/29) of samples, and TPI in 10.3 % (3/29) of samples. Only one sample from a dog showed the amplification of all genes. PCR-RFLP analysis showed assemblage F in a human, dog, and cat samples; and assemblage C and D in dog samples. This is the first description of assemblage F in humans from Brazil and the first description of assemblage F in dogs. Further studies are needed to verify the frequency with which these infections occur, and provide information that will contribute to the molecular epidemiological understanding of giardiasis.
Giardiasis
2022, Greene's Infectious Diseases of the Dog and Cat, Fifth Edition
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  Cause: Giardia duodenalis (multiple genetic assemblages); a flagellate protozoan (phylum Sarcomastigophora).
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  First Described: Trophozoites that were likely Giardia spp. were first described in 1681 by Antonie van Leeuwenhoek and the genus was established in the early 1900s. Infections of dogs and cats were first described in the 1960s.¹^,²
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  Affected Host Species: Dogs and cats are the definitive hosts for several different Giardia assemblages and pass either trophozoites (diarrhea) or cysts (diarrhea or normal stool) in feces.
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  Intermediate Hosts: Although there are no intermediate hosts, there are multiple potential transport hosts that may carry Giardia spp. cysts and indirectly infect dogs or cats.
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  Route of transmission: Fecal-oral.
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  Geographic Distribution: Worldwide.
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  Major Clinical Signs: Most dogs and cats harbor subclinical infections. Acute watery diarrhea that can contain mucus can occur. Steatorrhea can be observed. Chronic or intermittent diarrhea and weight loss may be noted in animals with concurrent infections or immunocompromise.
- •
  Differential Diagnoses: All causes of small bowel diarrhea including Cystoisospora spp., Cryptosporidium spp., bacterial infections such as salmonellosis, nematode infections, dietary intolerance, exocrine pancreatic insufficiency, and inflammatory bowel disease.
- •
  Human Health Significance: Giardia duodenalis assemblages that infect dogs (assemblages C and D) or cats (assemblage F) are usually not associated with clinical disease in humans but can be detected in the feces of immunocompromised people.³ Human assemblages A and B can be found in feces of dogs and cats when animals share human environmental features (such as contaminated water). However, it is unlikely that new human infections with G. duodenalis assemblage A or B infections are acquired from the feces of dogs or cats.
The controversies surrounding Giardia intestinalis assemblages A and B
2021, Current Research in Parasitology and Vector-Borne Diseases
Giardia intestinalis continues to be one of the most encountered parasitic diseases around the world. Although more frequently detected in developing countries, Giardia infections nonetheless pose significant public health problems in developed countries as well. Molecular characterisation of Giardia isolates from humans and animals reveals that there are two genetically different assemblages (known as assemblage A and B) that cause human infections. However, the current molecular assays used to genotype G. intestinalis isolates are quite controversial. This is in part due to a complex phenomenon where assemblages are incorrectly typed and underreported depending on which targeted locus is sequenced. In this review, we outline current knowledge based on molecular epidemiological studies and raise questions as to the reliability of current genotyping assays and a lack of a globally accepted method. Additionally, we discuss the clinical symptoms caused by G. intestinalis infection and how these symptoms vary depending on the assemblage infecting an individual. We also introduce the host-parasite factors that play a role in the subsequent clinical presentation of an infected person, and explore which assemblages are most seen globally.
Multilocus genotyping of Giardia intestinalis in pet dogs of Medellín Colombia
2021, Veterinary Parasitology: Regional Studies and Reports
Citation Excerpt :
Recently, in a Giardia prevalence study in children and eight dogs in southwest Colombia (city of Popayán), a single dog Giardia-positive sample was analyzed and genotyped as sub-assemblage BIII (Villamizar et al., 2019). Similar to our study, in Brazil, genotypes C and D were found in naturally infected dogs from Sao Paulo (Souza et al., 2007). Likewise, a study in Vietnam found infected dogs with Giardia belonging to assemblages C and D exclusively (Nguyen et al., 2018), as well as studies from Korea, Israel and Poland (Kim et al., 2019; Salant et al., 2020; Piekara-Stępińska et al., 2020).
According to a few parasitological and epidemiological studies, Giardia is the most prevalent parasitic infection among pet dogs in the city of Medellín, the second-largest city in Colombia. This study determined the assemblages of Giardia in the fecal samples of dogs obtained from 18 veterinary centers of Medellín. One hundred fecal samples of dogs diagnosed with Giardia using microscopy were analyzed via nested polymerase chain reaction (PCR) using three genes (gdh, bg, and tpi). The PCR products were purified and sequenced, and phylogenetic analyses were conducted using the maximum likelihood algorithm of the three loci. From the 100 samples analyzed, 47 were Giardia-positive via PCR. Genotypes C and D were detected in six samples, neither of which were associated with human infection. However, the zoonotic potential of Giardia cannot be ruled out because of the small number of samples that could be sequenced for assemblage assignation.
First description of Giardia duodenalis in buffalo calves (Bubalus bubalis) in southwest region of São Paulo State, Brazil
2019, Food and Waterborne Parasitology
We performed molecular characterization of Giardia duodenalis in buffalo calves from the Southwest region of São Paulo State, Brazil. A total of 183 fecal samples of Murrah breed buffaloes up to six months of age were collected. We examined these samples by the polymerase chain reaction (PCR) targeting the small-subunit ribosomal RNA gene and positive samples were characterized using additional PCR assays targeting a portion of the beta-giardin, the glutamate dehydrogenase and the triose-phosphate isomerase genes. Based on the SSU rRNA nPCR, the presence of G. duodenalis was confirmed in 12 (6.56%) of fecal samples, of these, five, four and three samples were positive for the tpi, bg and gdh genes, respectively. Assemblage identification by sequencing was successful in 6 of 12 samples and sequence analysis showed 100% genetic similarity with G. duodenalis assemblage E. This observation represents the first detection of G. duodenalis assemblage E in buffaloes calves in Brazil.

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Short communicationMolecular identification of Giardia duodenalis isolates from humans, dogs, cats and cattle from the state of São Paulo, Brazil, by sequence analysis of fragments of glutamate dehydrogenase (gdh) coding gene

Abstract

Section snippets

Acknowledgements

Vet. J.

Int. J. Parasitol.

Infect. Genet. Evol.

Infect. Genet. Evol.

Vet. Parasitol.

Vet. Parasitol.

Int. J. Parasitol.

Diagn. Microbiol. Infect. Dis.

Vet. Parasitol.

Adv. Parasitol.

Int. J. Parasitol.

Acta Trop.

J. Biol. Chem.

Identification of genotypes of Giardia intestinalis isolates from dogs in Japan by direct sequencing of the PCR amplified glutamate dehydrogenase gene

J. Vet. Med. Sci.

Short communication
Molecular identification of Giardia duodenalis isolates from humans, dogs, cats and cattle from the state of São Paulo, Brazil, by sequence analysis of fragments of glutamate dehydrogenase (gdh) coding gene