Intestinalis Isolates Research Articles

Gut microbial genomes with paired isolates from China illustrate probiotic and cardiometabolic effects

The gut microbiome displays genetic differences among populations, and characterization of the genomic landscape of the gut microbiome in China remains limited. Here, we present the Chinese Gut Microbial Reference (CGMR) set, comprising 101,060 high-quality metagenomic assembled genomes (MAGs) of 3,707 nonredundant species from 3,234 fecal samples across primarily rural Chinese locations, 1,376 live isolates mainly from lactic acid bacteria, and 987 novel species relative to worldwide databases. We observed region-specific coexisting MAGs and MAGs with probiotic and cardiometabolic functionalities. Preliminary mouse experiments suggest a probiotic effect of two Faecalibacillus intestinalis isolates in alleviating constipation, cardiometabolic influences of three Bacteroides fragilis_A isolates in obesity, and isolates from the genera Parabacteroides and Lactobacillus in host lipid metabolism. Our study expands the current microbial genomes with paired isolates and demonstrates potential host effects, contributing to the mechanistic understanding of host-microbe interactions.

Sequence and Haplotype Analyses of Ligula intestinalis in Acanthobrama marmid (Cyprinidae) in Turkey

PurposeLigulosis caused by Ligula intestinalis adversely affects the fisheries carried out in the lakes and ponds, causing economic losses in the fish industry. In this study, it was aimed to reveal the molecular characterization of L. intestinalis isolates obtained from woodfish (Acanthobrama marmid) in Keban Dam Lake in Elazig province of Turkey by using mt-CO1 gene sequences and to determine the genetic differences and haplotypes between the isolates.MethodsIn the examination made in terms of L. intestinalis, the intestine of the fish was opened with the help of fine-tipped scissors, the contents were allowed to come out, and the parasites were taken into a petri dish containing phosphate buffered saline (PBS). Then, L. intestinalis plerocercoids were taken into 15 ml falcon tubes containing 70% ethanol and stored at − 20 °C until further analysis. From each isolate, total gDNA was extracted from the plerocercoids. A partial (480 bp) mt-CO1 gene was amplified by PCR and sequenced unidirectionally. The final size of the trimmed sequences was 392 bp for 43 sequences. Sequence and haplotype analyses were performed, followed by phylogenetic analyses.ResultsAll isolates were confirmed as L. intestinalis by BLAST analysis. In addition, 87 nucleotide mutation positions were determined among 43 CO1 gene sequences. As a result of the haplotype network performed for the mt-CO1 gene region of L. intestinalis isolates; arranged in a star-like configuration with the main haplotype (Hap05), separated from other haplotypes by 1–6 mutation steps, and 29 haplotypes were identified, covering 13.9% (6/43) of the total isolates. Also, 75 variable (polymorphic) sites were determined, 52 of which were parsimony informative sites.ConclusionsThe molecular characterization of L. intestinalis in woodfish (A. marmid) was identified for the first time in Turkey.

Molecular appraisal of Giardia intestinalis from Western India: A prospective observational study.

Giardia intestinalis is an intestinal protozoan which commonly causes parasitic gastroenteritis globally. It is a species complex consisting of at least eight assemblages (genotypes). In India, Giardia is mostly underreported and missed in asymptomatic cases. The aim of this study was to genotype the G. intestinalis isolates from stool samples of patients at a tertiary care center in Rajasthan, India, and to clinically correlate it. This prospective pilot cross-sectional study was conducted from 2019 to 2021 in a tertiary care center in western India. Patients who were microscopically positive for giardiasis were enrolled. DNA was extracted from their stool samples and amplified by polymerase chain reaction (PCR) using 4E1-HP as the target sequence. Anthropometric measurements and analysis were done for children by using Anthrocal application. A total of 50 patients were enrolled. Diarrhea was present in 18 patients (36%). Among these, 6 were immunocompromised and had different comorbidities. Among the children <12 years of age, 55.17% (n = 16/29) were stunted (<-2 S.D.), and among <5 years, 44.4% (n = 4/9) showed wasting (<-2 S.D.). A PCR product corresponding to assemblage B of G. intestinalis was amplified in 47 stool specimens. Only three stool samples were negative for both assemblages A and B and posed an interesting enigma. In this study, a predominance of assemblage B of G. intestinalis was detected in 94% of the isolates. Furthermore, the possibility of zoonotic transmission could not be ruled out.

Characterization of Metronidazole-Resistant Giardia intestinalis Lines by Comparative Transcriptomics and Proteomics.

Metronidazole (MTZ) is a clinically important antimicrobial agent that is active against both bacterial and protozoan organisms. MTZ has been used extensively for more than 60 years and until now resistance has been rare. However, a recent and dramatic increase in the number of MTZ resistant bacteria and protozoa is of great concern since there are few alternative drugs with a similarly broad activity spectrum. To identify key factors and mechanisms underlying MTZ resistance, we utilized the protozoan parasite Giardia intestinalis, which is commonly treated with MTZ. We characterized two in vitro selected, metronidazole resistant parasite lines, as well as one revertant, by analyzing fitness aspects associated with increased drug resistance and transcriptomes and proteomes. We also conducted a meta-analysis using already existing data from additional resistant G. intestinalis isolates. The combined data suggest that in vitro generated MTZ resistance has a substantial fitness cost to the parasite, which may partly explain why resistance is not widespread despite decades of heavy use. Mechanistically, MTZ resistance in Giardia is multifactorial and associated with complex changes, yet a core set of pathways involving oxidoreductases, oxidative stress responses and DNA repair proteins, is central to MTZ resistance in both bacteria and protozoa.

Molecular and epidemiological characterization of Giardia Intestinalis assemblages detected in Djelfa, Algeria.

Giardia intestinalis is a flagellated protozoan that lives and proliferates in the small intestine of the host causing giardiasis. The route of transmission is the fecal-oral route, either directly or indirectly. Limited genetic information on G. intestinalis is known in Algeria. This study aimed to estimate the prevalence of G. intestinalis assemblages in the city of Djelfa. A total of 355 fecal samples were collected from symptomatic and asymptomatic school children aged ranged between 6 and 11years old. Genotyping was done to the Giardia positive samples (n = 30) targeting the beta-giardin gene by applying PCR/RFLP assay. Our data showed that most of the cases were asymptomatic (56.7%). Co-infection with other intestinal parasites was found in 16.6% of cases. We obtained 28/30 positive PCR products while two samples only showed false-negative results, and only 20 samples have shown strong PCR products suitable for RFLP analysis. Assemblage A (70%) was more prevalent than assemblage B (30%) and was more expressed by signs than assemblage B. Moreover, only assemblage A was associated with close contacts with domestic animals and birds. In conclusion, this study gave the first molecular data on G. intestinalis isolates in the city of Djelfa. Further expanded studies using more genes and covering other cities in Algeria are mostly needed.

Detections of gastrointestinal parasites, including Giardia intestinalis and Cryptosporidium spp., in cattle of Banten province, Indonesia.

Gastrointestinal parasites can induce low productivity in livestock by causing acute or chronic enteritis. Veterinarians make great efforts to design rational and effective hygienic protocols for both the prevention and treatment of diarrhea. Although prevalences can vary depending on the examined areas or the ages of the hosts, and the methods used for detections, it is helpful to accumulate data across many areas to evaluate parasitic distribution. A coprological survey in cattle was conducted in Tangerang, Banten Province of Indonesia, in order to determine the prevalence of the parasites, including those of diarrhea-associated diseases. Furthermore, the risk of transmission of Giardia intestinalis and Cryptosporidium spp. to human was genetically analyzed. Gastrointestinal parasites were detected in 87 of 109 cattle samples, including 85 carrying Eimeria spp., 36 carrying Fasciola gigantica, 35 carrying Strongyloides spp., 33 carrying Paramphistomum spp., and 15 carrying Capillaria spp. Giardia intestinalis and Cryptosporidium spp., parasites with zoonotic potential, were detected in 9 and 1 cattle samples, respectively. Molecular analyses identified the G. intestinalis isolate as a member of Assemblage E, which has been recently detected in humans in another country. These results may be helpful in understanding the hygienic risk affecting the livestock productivity and zoonotic potential of cattle in Indonesia.

Multi-locus genotyping and phylogenetic analyses of Giardia intestinalis isolates from indigenous goats in Ogun State, Nigeria

Giardiasis is a common gastrointestinal disease of humans and various animal species worldwide. In this study, 302 stool samples were collected from West African Dwarf and Sokoto Red breeds of goats in Ogun State, Nigeria, and screened for Giardia intestinalis coproantigens using enzyme-linked immunosorbent assay (ELISA). The genotypes of G. intestinalis in faecal samples collected from 152 goats raised on selected farms were identified by polymerase chain reaction (PCR) amplification and sequence analyses of the small subunit ribosomal RNA (ssu rRNA), glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi) and β-giardin (bg) genes. Based on ELISA, an overall prevalence of 45.7% was recorded with the infection rates in pre-weaned (60.2%) and post-weaned goat kids (51.5%) being significantly (p < 0.05) higher than in adults (28.2%). Giardia intestinalis DNA was amplified in 31.6% and 29.6% of goat faeces at the ssu rRNA and gdh loci respectively. These were genotyped at the ssu rRNA locus as assemblages B (n = 13) and E (n = 36). Similar results were observed at the gdh locus except that eight isolates contained assemblage E mixed with either assemblage A or B. Additionally, sub-assemblages BI (n = 7) and BIII (n = 2) were identified with up to four single nucleotide polymorphisms (SNPs) occurring in these isolates. Multilocus genotypes (MLG) of all assemblage E isolates were identified using the ssu rRNA and gdh loci while MLG of all isolates containing assemblage B and mixed assemblages were determined after further typing at the tpi and bg loci. Forty-two MLG isolates were identified and these comprised 32, 8 and 2 (sub)-assemblage E, BI and BIII respectively. All isolates with mixed assemblages at the gdh locus were consequently designated as assemblage E by MLG. The assemblage E isolates from goats were genetically related to isolates from cattle, sheep and goats while the assemblage B isolates were related to isolates of human, pig and lemur origin. This suggests that G. intestinalis isolated from goats bred in Ogun State, Nigeria may be capable of cross-species transmission and may be of zoonotic importance.

Association between enteric protozoan parasites and gastrointestinal illness among HIV- and tuberculosis-infected individuals in the Chowke district, southern Mozambique

Human immune deficiency virus (HIV) and tuberculosis (TB) infections remain major public health issues globally, particularly in sub-Saharan Africa. Impairment of both cell-mediated and humoral immunity by HIV and/or TB infections may limit the host’s defences against other pathogens, including the diarrheagenic protozoan Cryptosporidium spp., Giardia intestinalis, and Entamoeba histolytica. During September–December 2015 a cross-sectional study was conducted to assess the prevalence and molecular diversity of these enteric parasites among HIV- and/or TB-infected patients at a medical reference centre in Chowke district, southern Mozambique. A total of 99 stool specimens were initially screened by direct microscopy and further confirmed and characterised by molecular methods. DNA sequence analyses of the genes encoding the small subunit ribosomal RNA and the 60-kDa glycoprotein were used for the typing and sub-typing of Cryptosporidium isolates, respectively. G. intestinalis-positive isolates by real-time PCR were subsequently typed at the glutamate dehydrogenase locus. Differential diagnosis of E. histolytica/dispar was achieved by real-time PCR. G. intestinalis (8.1%) was the enteric protozoan more frequently detected, followed by Cryptosporidium spp. (7.1%), and Entamoeba histolytica/dispar (6.1%). Two HIV-infected (but not TB-infected) patients harbour G. intestinalis and Cryptosporidium spp. co-infections. Two (29%) G. intestinalis isolates were successfully characterised, revealing the presence of known AII and novel BIV genotypes. Four (57%) Cryptosporidium isolates were unmistakeable assigned to C. hominis, identifying two (IbA10G2 and IdA22) sub-types. Cryptosporidium infections were not associated to diarrhoea in HIV-positive patients, probably because improved immune function in the affected individuals due to antiretroviral therapy. G. intestinalis was considered a non-opportunistic pathogen, whereas the presence of E. histolytica could not be confirmed by molecular methods. Based on their common presence in the studied clinical population, we recommend the effective diagnosis and treatment of these enteropathogens for improving the management of HIV and TB patients.

Sequence variation in three mitochondrial genes among Gasterophilus intestinalis isolates from two distinct regions in China

Horse gastrointestinal myiasis caused by larvae of Gasterophilus spp. has a worldwide distribution. However, little information is available on the genetic variation of Gasterophilus spp. at the molecular level. In the present study, sequence variation was examined in three mitochondrial (mt) genes, namely mt cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunits 5 (nad5) and large subunit ribosomal RNA (rrnL), among G. intestinalis isolates originating from Heilongjiang Provience and Xinjiang Uygur Autonomous Region, China. The complete cox1, nad5, and rrnL gene sequences were amplified by PCR separately from adult G. intestinalis individuals and the amplicons were sequenced from both directions. The size of the sequences of cox1, nad5, and rrnL genes was 1539 bp, 1717 bp, and 1321 bp, respectively. The A + T contents of the sequences were 62.7–63.9% (cox1), 70–70.7% (nad5), and 74.5–74.9% (rrnL). The intra-specific sequence variations within G. intestinalis were 0.1–3.1% for cox1, 0.4–3.4% for nad5, and 0.1–1.5% for rrnL. Phylogenetic analyses based on the cox1 sequences using the Bayesian inference indicated that all the G. intestinalis isolates grouped together with high statistical support. These findings demonstrated clearly the usefulness of mt cox1, nad5, and rrnL sequences for the molecular identification and for studying population genetics of G. intestinalis in horses.

Bacteroides faecis and Bacteroides intestinalis recovered from clinical specimens of human intestinal origin.

We report three cases of recently named Bacteroides spp. isolates, two B. faecis isolates and one B. intestinalis isolate from clinical specimens of inpatients at a Korean tertiary-care hospital in 2011. All isolates were susceptible to piperacillin-tazobactam, imipenem, meropenem, chloramphenicol, and metronidazole.

A low-virulence Eimeria intestinalis isolate from rabbit (Oryctolagus cuniculus) in China: molecular identification, pathogenicity, and immunogenicity

An Eimeria intestinalis isolated from a rabbit in China was first identified by amplifying the 18S small subunit (SSU) ribosomal RNA gene. The size of the amplified fragment was 1521 bp. The 18S SSU RNA gene of the E. intestinalis isolate shared 99% sequence identity with E. intestinalis isolates from France and the Czech Republic, with 100 and 96% coverage, respectively. Then, the pathogenicity and immunogenicity of the E. intestinalis isolate were evaluated in specific pathogen free (SPF) rabbits. In the pathogenicity assay, SPF rabbits in four groups were infected with 5 × 10(3), 5 × 10(4), 5 × 10(5), and 0 sporulated oocysts, respectively. Clinical signs including diarrhoea, constipation, loss of appetite, and reduction of body weight gain were observed in rabbits inoculated with 5 × 10(4) and 5 × 10(5) oocysts. And one rabbit (25 %) inoculated with 5 × 10(5) oocysts died 15 days after the inoculation. In the immunogenicity assay, SPF rabbits in five groups (named B1, B2, B3, B4, and B5) were immunised with 5 × 10(1), 5 × 10(2), 5 × 10(3), 0, and 0 sporulated oocysts, respectively. All rabbits but the B5 group were challenged with 1 × 10(6) oocysts. After the challenge, no or slight clinical signs were seen in rabbits of the B2 and B3 groups. Compared with the control, a 69.6 and 84.5% reduction of oocyst output was observed in the B2 and B3 groups, respectively. The body weight gain of the two groups was obviously higher than that of the challenge control group. All the results show that the E. intestinalis isolate has low virulence but immunogenicity in rabbit.

The epidemiology of Giardia intestinalis assemblages A and B among Egyptian children with diarrhea: A PCR-RFLP-based approach

Background The protozoan parasite Giardia intestinalis is a common childhood infection in developing countries that causes diarrheal illness. The majority of G. intestinalis isolates from humans are grouped into two distinct genetic assemblages A and B. The molecular epidemiological studies on G. intestinalis assemblages in humans are limited in Egypt. Objective This study was conducted to estimate the detection rate of G. intestinalis infection among a cohort of children suffering from diarrhea in the Dakahalia governorate, Egypt, and to correlate between clinical giardiasis and Giardia spp. assemblages in positive stool samples by PCR-restriction fragment length polymorphism (PCR-RFLP). Participants and methods A total of 311 diarrheal stool samples were examined microscopically for Giardia spp. infection. DNA samples were isolated from the stools of 103 (33.12%) positive samples with G. intestinalis , amplified with PCR, and digested with the Xho I enzyme for RFLP. Results Of the 103 samples, 64 (62.14%) were found to be assemblage B, whereas 32 samples (31.07%) belonged to assemblage A. Mixed genotype A and B was present in three samples (2.91%), and four samples (3.88%) were of undetermined Giardia spp. assemblage. The detection rate of assemblage B was higher in samples from children with persistent diarrhea, whereas assemblage A detection rate was higher in samples from acute diarrhea. Conclusion G. intestinalis causing diarrhea in children in the Dakahalia governorate, Egypt, predominantly belongs to assemblage B, indicating that human-to-human method of infection is more common than zoonotic method.

Molecular characterisation of Giardia intestinalis assemblages from human isolates at a tertiary care centre of India

Purpose: The aim of the study was to determine the genetic heterogeneity of Giardia intestinalis isolates detected in stool samples of the study population using polymerase chain reaction assay and restriction fragment length polymorphism. We also tried to correlate the association/differences between the clinical symptomatology and infection by different assemblages (genotypes) of G. intestinalis. Materials and Methods: This cross-sectional study was conducted from April 2008 to June 2010. A total of 40 adults (n = 40) and 42 children (n = 42) below the age of 12 years with the clinical suspicion of giardiasis and with the onset of one or more of the following five symptoms, i.e., loose stool, nausea, weight loss, fatigue and foul smelling faeces and confirmed laboratory diagnosis of giardiasis at least once during the current episode of diarrhoea were included in this study. Results: Of the 82 patients (males 66) enrolled in the study, 70 (85%) presented with diarrhoea (56 males) and 12 (15%) without diarrhoea (10 males). Out of 70 diarrheic patients, 61 (87%) had chronic diarrhoea, 8 (11.5%) had acute diarrhoea and 1 (1.5%) had persistent diarrhoea. Of the total patients, 63 (77%) were clinically assessed and were apparently immunocompetent, whereas, 19 (23%) immunocompromised patients had different underlying conditions besides giardiasis. Genotyping identified all 82 (100%) isolates as assemblage B. Conclusion: We found that assemblage B of G. intestinalis presents with all kinds of clinical features ranging from asymptomatic carriage to acute, persistent or chronic diarrhoea.

Molecular investigation of zoonotic genotypes of Giardia intestinalis isolates in humans, dogs and cats, sheep, goats and cattle in Araçatuba (São Paulo State, Brazil) by the analysis of ß-giardin gene fragments

In the period from July 2009 to October 2010, fecal samples from 61 animals and 154 humans from the municipality of Aracatuba (Sao Paulo State, Brazil) were studied. Fecal samples from animals were collected in the Municipal Animal Shelter and the Veterinary Hospital of the Universidade Estadual Paulista. Human fecal specimens were collected in playschools in the outskirts of the city by the private network of clinical analysis laboratories of the municipal. Diagnosis was done by optical microscopy using the Faust and Hoffmann, Pons and Janer techniques. The genotypes of Giardia intestinalis were characterized by PCR-RFLP and confirmed by sequencing the s-giardin gene. Human specimens were positive in 25.3% (39/154) of the cases with 26.8% (36/134) of the specimens from children and 15% (3/20) from adults being positive. The frequency of G. intestinalis among the animals was 23.0% (14/61). A total of 32 isolates of G. intestinalis obtained from human feces and six from dogs and cats were characteristic of the A genotype (AI and AII/AIII). The results of this study in respect to frequency of giardiasis are similar to reported in most studies in Brazil. The prevalence observed in animal populations conforms to worldwide infection rates. G. intestinalis genotypes considered zoonotic were detected in both pets and humans from the city of Aractuba, suggesting a possible zoonotic transmission of the parasite in the northwestern region of Sao Paulo State. The absence of these genotypes in farm animals may imply that they are not involved in the chain of transmission to humans in this region.

Arginine Consumption by the Intestinal Parasite Giardia intestinalis Reduces Proliferation of Intestinal Epithelial Cells

In the field of infectious diseases the multifaceted amino acid arginine has reached special attention as substrate for the host´s production of the antimicrobial agent nitric oxide (NO). A variety of infectious organisms interfere with this part of the host immune response by reducing the availability of arginine. This prompted us to further investigate additional roles of arginine during pathogen infections. As a model we used the intestinal parasite Giardia intestinalis that actively consumes arginine as main energy source and secretes an arginine-consuming enzyme, arginine deiminase (ADI). Reduced intestinal epithelial cell (IEC) proliferation is a common theme during bacterial and viral intestinal infections, but it has never been connected to arginine-consumption. Our specific question was thereby, whether the arginine-consumption by Giardia leads to reduced IEC proliferation, in addition to NO reduction. In vitro cultivation of human IEC lines in arginine-free or arginine/citrulline-complemented medium, as well as in interaction with different G. intestinalis isolates, were used to study effects on host cell replication by MTT assay. IEC proliferation was further analyzed by DNA content analysis, polyamine measurements and expressional analysis of cell cycle regulatory genes. IEC proliferation was reduced upon arginine-withdrawal and also in an arginine-dependent manner upon interaction with G. intestinalis or addition of Giardia ADI. We show that arginine-withdrawal by intestinal pathogens leads to a halt in the cell cycle in IECs through reduced polyamine levels and upregulated cell cycle inhibitory genes. This is of importance with regards to intestinal tissue homeostasis that is affected through reduced cell proliferation. Thus, the slower epithelial cell turnover helps the pathogen to maintain a more stable niche for colonization. This study also shows why supplementation therapy of diarrhea patients with arginine/citrulline is helpful and that citrulline especially should gain further attention in future treatment strategies.

Identification of a novel Assemblage B subgenotype and a zoonotic Assemblage C in human isolates of Giardia intestinalis in Egypt

Giardia intestinalis ( G. intestinalis) is a flagellate parasite which has been considered the most common protozoan infecting human. Molecular techniques are of great value in studying the taxonomy, the zoonotic potential of animal isolates and the correlation between the genetic variability of the parasite and the range of clinical symptoms observed in humans. The present work aims at genotyping G. intestinalis isolates from Egypt using molecular techniques. PCR targeting the β-giardin locus, RFLP and sequencing were applied to 12 microscopically positive and 3 microscopically negative samples (which were positive by real time PCR targeting SSUr DNA). Two other loci, triose phosphate isomerase (TPI) gene and glutamate dehydrogenase (GDH) gene PCR and RFLP were also applied to all study isolates. The most frequent genotype was Assemblage B (13 out of 15), while Assemblage A and C were present in one sample each. This is the first report on zoonotic transmission of Assemblage C (dog genotype) to human in Egypt. Sequencing of the Assemblage B isolates revealed new subgenotypes with consistent mutations at specific positions, some of which were not characterized previously. The results shed light on the possibility that G. intestinalis can infect humans through a zoonotic route and open the door to wider investigations using different genetic loci to genotype Giardia isolates.

Multilocus Genotyping of Human Giardia Isolates Suggests Limited Zoonotic Transmission and Association between Assemblage B and Flatulence in Children

Background Giardia intestinalis is one of the most common diarrhea-related parasites in humans, where infection ranges from asymptomatic to acute or chronic disease. G. intestinalis consists of eight genetically distinct genotypes or assemblages, designated A–H, and assemblages A and B can infect humans. Giardiasis has been classified as a possible zoonotic disease but the role of animals in human disease transmission still needs to be proven. We tried to link different assemblages and sub-assemblages of G. intestinalis isolates from Swedish human patients to clinical symptoms and zoonotic transmission.Methodology/Principal FindingsMultilocus sequence-based genotyping of 207 human Giardia isolates using three gene loci: ß-giardin, glutamate dehydrogenase (gdh), and triose phosphate isomerase (tpi) was combined with assemblage-specific tpi PCRs. This analysis identified 73 patients infected with assemblage A, 128 with assemblage B, and six with mixed assemblages A+B. Multilocus genotypes (MLGs) were easily determined for the assemblage A isolates, and most patients with this genotype had apparently been infected through anthroponotic transmission. However, we also found evidence of limited zoonotic transmission of Giardia in Sweden, since a few domestic human infections involved the same assemblage A MLGs previously reported in Swedish cats and ruminants. Assemblage B was detected more frequently than assemblage A and it was also more common in patients with suspected treatment failure. However, a large genetic variability made determination of assemblage B MLGs problematic. Correlation between symptoms and assemblages was found only for flatulence, which was significantly more common in children less than six years of age infected with assemblage B.Conclusions/SignificanceThis study shows that certain assemblage A subtypes are potentially zoonotic and that flatulence is connected to assemblage B infections in young children. Determination of MLGs from assemblages A and B can be a valuable tool in outbreak situations and to help identify possible zoonotic transmission.

Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan

The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP ® Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase ( gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan.

Genome analysis and comparative genomics of a Giardia intestinalis assemblage E isolate

BackgroundGiardia intestinalis is a protozoan parasite that causes diarrhea in a wide range of mammalian species. To further understand the genetic diversity between the Giardia intestinalis species, we have performed genome sequencing and analysis of a wild-type Giardia intestinalis sample from the assemblage E group, isolated from a pig.ResultsWe identified 5012 protein coding genes, the majority of which are conserved compared to the previously sequenced genomes of the WB and GS strains in terms of microsynteny and sequence identity. Despite this, there is an unexpectedly large number of chromosomal rearrangements and several smaller structural changes that are present in all chromosomes. Novel members of the VSP, NEK Kinase and HCMP gene families were identified, which may reveal possible mechanisms for host specificity and new avenues for antigenic variation. We used comparative genomics of the three diverse Giardia intestinalis isolates P15, GS and WB to define a core proteome for this species complex and to identify lineage-specific genes. Extensive analyses of polymorphisms in the core proteome of Giardia revealed differential rates of divergence among cellular processes.ConclusionsOur results indicate that despite a well conserved core of genes there is significant genome variation between Giardia isolates, both in terms of gene content, gene polymorphisms, structural chromosomal variations and surface molecule repertoires. This study improves the annotation of the Giardia genomes and enables the identification of functionally important variation.

From mouse to moose: Multilocus genotyping of Giardia isolates from various animal species

Giardia intestinalis is a protozoan parasite that consists of seven genetically distinct assemblages (A to G). Assemblage A and B parasites have been detected in a wide range of animals including humans, while the other assemblages (C to G) appear to have a narrower host range. However, the knowledge about zoonotic transmission of G. intestinalis is limited. To address this question, 114 Giardia isolates from various animals in Sweden including pets, livestock, wildlife and captive non-human primates were investigated by a sequence-based analysis of three genes (β-giardin, glutamate dehydrogenase and triose phosphate isomerase). Assemblage A infections were detected in nine ruminants, five cats and one dog, while three sheep were infected with both assemblages A and E. Multilocus genotypes (MLGs) were defined for assemblage A, and three of these MLGs have previously been detected in Giardia isolates from humans. The newly described sub-assemblage AIII, until now reported mainly in wild hoofed animals, was found in one cat isolate. Assemblage B occurred in three monkeys, one guinea pig and one rabbit. The rabbit isolate exhibited sequences at all three loci previously detected in human isolates. The non-zoonotic assemblages C, D, E, F or G were found in the remaining 83 G. intestinalis isolates, which were successfully amplified and genotyped, generating a wide variety of both novel and known sub-genotypes. Double peaks in chromatograms were seen in assemblage B, C, D and E isolates but were never observed in assemblage A, F and G isolates, which can reflect differences in allelic sequence divergence. No evidence of genetic exchange between assemblages was detected. The study shows that multilocus genotyping of G. intestinalis is a highly discriminatory and useful tool in the determination of zoonotic sub-groups within assemblage A, but less valuable for subtyping assemblages B, C, D and E due to the high frequency of double peaks in the chromatograms. The obtained data also suggest that zoonotic transmission of assemblages A and B might occur to a limited extent in Sweden.