Giardia Genotypes Research Articles

Genotyping and epidemiological distribution of diarrhea-causing isolates of Giardia duodenalis in southeastern part of West Bengal, India.

The prevalence and genetic diversity of the protozoan pathogen Giardia duodenalis have been extensively studied worldwide. There is currently a lack of data regarding the genetic variability of the organism in eastern India. Understanding the circulating genotypes and associated risk factors is crucial for effective planning and implementing control measures. Therefore, the objective of the study was to conduct an epidemiological study to determine the prevalence and identify the various genotypes present. This survey adds to our knowledge on the occurrence and distribution of Giardia genotypes in the studied region. The overall prevalence was found to be 6.8%. This parasitic infection was significantly associated with two age groups, i.e., >0-5 years and >5-12 years. Using a multilocus genotyping method, we genotyped 52 human Giardia isolates that were obtained from diarrheal patients. Two distinct assemblages were found in the population-30.8% belonged to assemblage A; 63.5% belonged to assemblage B, prevalent in the population; and 5.7% belonged to a combined assemblage A+B. Sub-assemblage AII was found in 17.3% of the cases, followed by sub-assemblage AI (13.5%). High levels of genetic diversity were found within the population of assemblage B undergoing balancing selection. Overall, the high prevalence of the parasite observed, particularly among children, raises a major concern and necessitates implementation of robust control measures. Furthermore, we report the presence of numerous unique genotypes, circulating in this limited geographical boundary, which can be useful dataset for future studies.

Molecular characterization of Cryptosporidium and Giardia in environmental samples and faecal samples from biogas users in Bac Giang, Vietnam

Little is known about Cryptosporidium and Giardia in biogas waste and humans in Vietnam. There is a potential risk of infections during or after using the biogas system. The detected protozoan genotypes are zoonotic pathogens, and contamination of vegetables may relay through runoff to the surface waters and soil. The objective of this study was to understand the role of the environment in the epidemiology of human infections in Bac Giang province, Vietnam, with a focus on investigating the presence of Cryptosporidium spp. genotypes and Giardia assemblages among 239 environmental samples and 94 faecal samples of biogas users. PCR and sequencing analysis were used to identify the occurrence and genotypes of Cryptosporidium and Giardia in these samples. Results showed that 13/333 (3.9 %) and 9/333 (2.7 %) samples were positive for Cryptosporidium oocyst and Giardia cysts, respectively. Characterization revealed the presence of Cryptosporidium scrofarum, C. suis, C. meleagridis, C. bailey and Giardia intestinalis assemblage A and E. C. scrofarum and Giardia assemblage E were identified for the first time in humans in Bac Giang. The current information from the above investigations will be valuable for protozoan source tracking and control interventions against Cryptosporidium and Giardia infection associated with biogas wastes in Vietnam.

Possible Correlation between Giardia duodenalis Genotypes and Fecal Calprotectin in Children with Diarrhea.

Giardiasis is one of the leading causes of diarrhea, particularly among children under the age of five in developing countries. Fecal calprotectin (FC) is an important biomarker for diagnosis and monitoring of inflammtory bowel disease, but other diagnoses should be considered in light of its elevation. We aimed to evaluate FC level in patients diagnosed with giardiasis and elucidate a possible correlation between Giardia genotypes and FC levels. Overall, 120 fecal samples were collected from children aged 4-12 years and tested for giardiasis by light microscopy. Out of which, 50 samples were enrolled within two groups: group I "cases" and group II "controls" and then subjected to PCR amplification, sequencing of the beta-giardin (bg) gene of the parasite, and FC evaluation. Assemblage B was identified in 75%, and assemblage A in 25% of samples. FC levels were statistically elevated in "group I" in comparison to "group II". Likewise, there was a statistically significant difference between FC levels in patients infected with assemblage A and assemblage B with a mean of 114 μg/gm and 202 μg/gm, respectively. The study highlighted the possible association between Giardia genotype B and elevated FC levels, further detailed studies are necessary to clarify these finding.

Zoonotic Giardia duodenalis Genotypes and Other Gastrointestinal Parasites in a Badger Population Living in an Anthropized Area of Central Italy.

The Eurasian badger (Meles meles) is widespread in Italy and occupies different habitats. The occurrence and species of gastrointestinal parasites were evaluated in a free-ranging badger population living in a highly anthropic area in central Italy. A total of 43 fecal samples were examined using the flotation test, the Mini-FLOTAC and Baermann techniques, and a rapid immunoassay for the detection of Giardia duodenalis and Cryptosporidium spp. fecal antigens. Molecular investigations were also performed that aimed at identifying Giardia genotypes. Overall, 37/43 samples (86%) were found positive. Specifically, 48.8% (21 samples) were positive for G. duodenalis, 23.2% (10/43) for Cryptosporidium spp., and 7% (3/43) for coccidian oocysts. Strongyloides sp. nematode larvae were detected in 3/43 samples (7%). Ascarid (1/43, 2.3%), capillariid (1/43, 2.3%), and strongyle-type eggs (76.7%, 33/43) were also identified. Among the 11 readable sequences of samples that were positive for G. duodenalis by end-point PCR (18/21), the zoonotic assemblage A sub-assemblage AII and mixed assemblage A and B were identified. This is the first report of zoonotic G. duodenalis genotypes in the Eurasian badger. Moreover, most of identified parasites have zoonotic potential and/or potential impact on the population health of wild badgers and other wild and domestic animals.

Humans and Hoofed Livestock Are the Main Sources of Fecal Contamination of Rivers Used for Crop Irrigation: A Microbial Source Tracking Approach.

Freshwater bodies receive waste, feces, and fecal microorganisms from agricultural, urban, and natural activities. In this study, the probable sources of fecal contamination were determined. Also, antibiotic resistant bacteria (ARB) were detected in the two main rivers of central Chile. Surface water samples were collected from 12 sampling sites in the Maipo (n = 8) and Maule Rivers (n = 4) every 3 months, from August 2017 until April 2019. To determine the fecal contamination level, fecal coliforms were quantified using the most probable number (MPN) method and the source of fecal contamination was determined by Microbial Source Tracking (MST) using the Cryptosporidium and Giardia genotyping method. Separately, to determine if antimicrobial resistance bacteria (AMB) were present in the rivers, Escherichia coli and environmental bacteria were isolated, and the antibiotic susceptibility profile was determined. Fecal coliform levels in the Maule and Maipo Rivers ranged between 1 and 130 MPN/100-ml, and 2 and 30,000 MPN/100-ml, respectively. Based on the MST results using Cryptosporidium and Giardia host-specific species, human, cattle, birds, and/or dogs hosts were the probable sources of fecal contamination in both rivers, with human and cattle host-specific species being more frequently detected. Conditional tree analysis indicated that coliform levels were significantly associated with the river system (Maipo versus Maule), land use, and season. Fecal coliform levels were significantly (p < 0.006) higher at urban and agricultural sites than at sites immediately downstream of treatment centers, livestock areas, or natural areas. Three out of eight (37.5%) E. coli isolates presented a multidrug-resistance (MDR) phenotype. Similarly, 6.6% (117/1768) and 5.1% (44/863) of environmental isolates, in Maipo and Maule River showed and MDR phenotype. Efforts to reduce fecal discharge into these rivers should thus focus on agriculture and urban land uses as these areas were contributing the most and more frequently to fecal contamination into the rivers, while human and cattle fecal discharges were identified as the most likely source of this fecal contamination by the MST approach. This information can be used to design better mitigation strategies, thereby reducing the burden of waterborne diseases and AMR in Central Chile.

Association Between Giardia Genotype and Oxidative Stress Biomarkers Among Giardia-Infected Children: A Case–Control Study

PurposeGiardia duodenalis is the most common worldwide intestinal protozoal infection. The implication of free radicals in organ injury occurs through oxidative stress. Infections as Giardia may act as a triggering or promoting factor for oxidative stress, particularly in children with compromised immunity. Besides, the effect of Giardia genotype on oxidative stress status is yet to be explored. Therefore, we sought to compare the oxidative stress status between Giardia positive cases (case group) and Giardia negative cases (control group), and to explore the association between Giardia genotype and the level of oxidative stress markers in Giardia-infected children, especially those receiving immunosuppressive therapy.MethodsPediatric patients attending Mansoura University Children Hospital in the period from April 2015 to October 2016 were enrolled. Both case (n = 50) and control (n = 50) groups were further subdivided into immunosuppressive therapy recipients (ITR) and non-immunosuppressive therapy recipients (NITR). Genotyping of Giardia from positive stool samples by PCR was carried out, and oxidative stress markers were measured from venous blood samples.ResultsGiardia positive cases had higher levels of Malondialdehyde (MDA) and lower levels of total antioxidant capacity (TAC). MDA highest level was associated with mixed genotypes A and B, while the highest TAC level was associated with Giardia genotype A in both ITR and NITR cases.ConclusionElevated oxidative stress biomarkers in pediatric patients infected with specific Giardia genotypes should receive considerable attention, because if prompt treatment is not conducted, oxidative damage may occur in patients with giardiasis, especially those receiving immunosuppressive therapy.

GIARDIA DUODENALIS GENOTYPING NOT LINKED TO CLINICAL SYMPTOMATOLOGY AND NUTRITIONAL STATUS OF SCHOOL-AGED CHILDREN OF SOLEDAD AND GALAPA MUNICIPALITY SCHOOLS, ATLÁNTICO, COLOMBIA.

Giardia duodenalis genotypes A and B have been reported in Colombia. The population consisted of 235 schoolchildren whose ages ranged from 2 to 10 yr of age from the municipalities of Soledad and Galapa in the department of Atlántico, Colombia. Fecal samples were obtained and then analyzed in triplicate using the sedimentation in formalin-ether (Ritchie's method) and direct examination techniques. Of the 235 fecal samples, 35 samples were positive for G. duodenalis; positive samples were concentrated in a sucrose gradient and sonicated for 3 cycles of 20 sec. DNA extraction was performed, and the parasites were genotyped by conventional PCR amplifying a region of the β-giardin gene. A general prevalence of G. duodenalis of 13.2% was found, and of these genotyped samples, 13 (56.7%) and 7 (20%) corresponded to genotype A, 1 (4.3%), and 3 (25%) corresponded to genotype B, and 9 (39.1%) and 2 (16.7%) were not defined, in the municipalities Soledad and Galapa, respectively. Additionally, 23 children were diagnosed with symptomatologic giardiasis, and 12 were asymptomatic; the most relevant symptoms were abdominal pain (7, 20%) and diarrhea (13, 56.7%). The nutritional status of children with Giardia genotypes A and B were as follows: 3 in a state of malnutrition (10%), 10 normal (33.3%), and 6 overweight and obese (20%) with genotype A, and 1 in a state of malnutrition (3.3%) and 3 normal (10%) with genotype B. The genotypes found in G. duodenalis did not show an association with nutritional status or with the clinical manifestations evaluated in schoolchildren.

Molecular characterisation of Giardia duodenalis from human and companion animal sources in the United Kingdom using an improved triosephosphate isomerase molecular marker

Giardia duodenalis is a protozoan parasite known for its ability to cause gastrointestinal disease in human and non-human mammals. In the UK, the full impact of this parasite has yet to be fully explored, due to the limited testing which has been undertaken in humans and the low-resolution assemblage-typing methods currently available. Rather than being primarily a travel-associated condition, a recent study has highlighted that an endemic Giardia cycle is present in the UK, although the source of human disease is unclear in the majority of cases. This study focussed on the improvement of one of the commonly used assemblage-typing assays, a nested topoisomerase phosphate (tpi) PCR, to increase the amplification success rate across both human and companion animal samples. After comparing published primers to full Giardia reference genomes, this marker protocol was optimised and then deployed to test a substantial number of human (n ​= ​79) and companion animal (n ​= ​174) samples to gain an insight into the molecular epidemiology of Giardia in the UK. One assemblage A1 and eleven assemblage A2 genotypes were detected in humans, along with and 25 assemblage B genotypes. Assemblage A1 genotypes, known to be human-infective, were found in three feline and one canine sample, while one feline sample contained assemblage A2. Additionally, four feline samples contained assemblage B, which is recognised as potentially human-infective. This study demonstrates the presence of potentially human-infective Giardia genotypes circulating in the companion animal population, notably with 17.4% (8/46) of feline-derived Giardia strains being potentially zoonotic. Using a modified tpi-based genotyping assay, this work highlights the potential for domestic pets to be involved in the endemic transmission of giardiasis in the UK and underlines the need for appropriate hygiene measures to be observed when interacting with both symptomatic and asymptomatic animals. It also serves to underline the requirement for further studies to assess the zoonotic risk of Giardia associated with companion animals in high-income countries.

Detection of Cryptosporidium spp. and Giardia duodenalis in small wild mammals in northeastern Brazil.

This study investigated the occurrence of Giardia duodenalis and Cryptosporidium spp. in rodents and marsupials from the Atlantic Forest in southern Bahia, northeastern Brazil. Two hundred and four fecal samples were collected from different forest areas in the municipalities of Ilhéus, Una, Belmonte, and Mascote. Identifications were performed using PCR and nested PCR followed by sequencing of the gdh and tpi genes for G. duodenalis, and the gp60 and Hsp-70 genes for Cryptosporidium. The total frequency of positive PCR samples for both G. duodenalis and Cryptosporidium spp. was 5.4% (11/204). Giardia duodenalis occurred in 2.94% (4/136) of rodents and 2.94% (2/68) of marsupials. The prevalence of Cryptosporidium in rodents and marsupials was 1.47% (2/136) and 4.41% (3/68), respectively. In the areas sampled, the frequency of parasitism was 50% (7/14), while the Mascote region alone had no parasitized animals. The G. duodenalis subgenotype AI was identified in the rodent species Hylaeamys laticeps, Oecomys catherinae, Oligoryzomys nigripes and Akodon cursor, and in the marsupials Gracilinanus agilis and Monodelphis americana. In the rodents Rhipidomys mastacalis, H. laticeps and in the marsupial Marmosa murina the protozoa Cryptosporidium fayeri, Cryptosporidium parvum and Cryptosporidium ubiquitum with subtypes IIa and IVg by the gp60 gene were found. In conclusion, this study provides the genetic characterization of Giardia and Cryptosporidium species and genotypes in rodents and marsupials. And, these findings reinforce that the rodent and marsupial species mentioned above play a role as new hosts for Giardia and Cryptosporidium.

Real-Time PCR for Molecular Detection of Zoonotic and Non-Zoonotic Giardia spp. in Wild Rodents

Giardiasis in humans is a gastrointestinal disease transmitted by the potentially zoonotic Giardia duodenalis genotypes (assemblages) A and B. Small wild rodents such as mice and voles are discussed as potential reservoirs for G. duodenalis but are predominantly populated by the two rodent species Giardia microti and Giardia muris. Currently, the detection of zoonotic and non-zoonotic Giardia species and genotypes in these animals relies on cumbersome PCR and sequencing approaches of genetic marker genes. This hampers the risk assessment of potential zoonotic Giardia transmissions by these animals. Here, we provide a workflow based on newly developed real-time PCR schemes targeting the small ribosomal RNA multi-copy gene locus to distinguish G. muris, G. microti and G. duodenalis infections. For the identification of potentially zoonotic G. duodenalis assemblage types A and B, an established protocol targeting the single-copy gene 4E1-HP was used. The assays were specific for the distinct Giardia species or genotypes and revealed an analytical sensitivity of approximately one or below genome equivalent for the multi-copy gene and of about 10 genome equivalents for the single-copy gene. Retesting a biobank of small rodent samples confirmed the specificity. It further identified the underlying Giardia species in four out of 11 samples that could not be typed before by PCR and sequencing. The newly developed workflow has the potential to facilitate the detection of potentially zoonotic and non-zoonotic Giardia species in wild rodents.

Giardiasis in urban and rural Amazonas, Brazil is driven by zoonotic and cosmopolitan A and B assemblages.

BACKGROUND Giardia duodenalis is a protozoan parasite that infects humans and other mammals and causes giardiasis worldwide. Giardia is genotyped into eight assemblages (A-H), with assemblages A and B considered zoonotic.OBJECTIVES The aim of this study was to determine the assemblages of G. duodenalis from individuals living in rural and urban areas of the Amazonas State.METHODS 103 human faecal specimens microscopically positive for the presence of Giardia obtained from four municipalities in Amazonas and four animal faecal specimens were genotyped based on the sequences of two genes, triosephosphate isomerase (TPI) and β-giardin (BG).FINDINGS In humans, assemblage A was the most represented with the identification of sub-assemblages AI, AII and AIII based on BG and sub-assemblages AI and AII based on TPI. Similarly, there is a diversity of sub-assemblage B considering BG (B and BIII) and TPI (B, BIII and BIV). In addition, we characterised homogeneous and heterogeneous genotypes comprising assemblages/sub-assemblages A and B in individuals from urban and rural areas. Here, for the first time, it was genotyped Giardia that infects animals from the Brazilian Amazon region. We identified sub-assemblage AI in one Ateles paniscus and two Felis catus and sub-assemblage BIV in one Lagothrix cana.MAIN CONCLUSIONS Therefore, humans and animals from the urban and rural Amazon share Giardia genotypes belonging to assemblages A and B, which are found in cosmopolitan regions around the world.

Asemptomatik köpeklerde görülen Giardia spp. enfeksiyonlarının zoonotik önemi

Giardia intestinalis (G. lamblia, G. duodenalis) is a common protozoan in human and other mammals worldwide causing diarrhea, vomiting, abdominal pain, weight loss and dehydration. The major infection occurs via directly faecal-oral route or indirectly contaminated food and water consumption. Dogs can carry the zoonotic genotypes of Giardia without any clinical signs and contaminate the environment. Asymptomatic carrier dogs living in the same environment with humans, food and water contaminated with these dogs' faeces can be a source of infection for humans. The aim of this review is to emphasize the asymptomatic course of Giardia spp. infections in dogs and to evaluate the effects of asymptomatic carriage on human health and the environment.

First multilocus sequence typing (MLST) of Giardia duodenalis isolates from humans in Romania

BackgroundGiardia duodenalis is one of the most prevalent and highly diverse human parasites, encompassing a complex of eight genetically distinct assemblages, each further divided into sub-assemblages. While in recent years, G. duodenalis genotype distribution patterns in humans have been intensely studied, there is still very little information available on the diversity of Giardia genotypes and sub-assemblages infecting people in Romania. In the present study, we investigated the genetic diversity of Giardia duodenalis in asymptomatic patients from Romania.MethodsOver an 11-month period, human feces from 7805 healthy adults were screened by microscopic analysis for G. duodenalis cysts during their obligatory periodic check-ups. DNA extraction was performed from microscopic-positive fecal samples, followed by multilocus sequence typing of four genetic loci of the ITS region, gdh, tpi and bg genes, followed by DNA sequencing and phylogenetic analysis. Statistical analysis was performed using EpiInfo 2000 software.ResultsThe prevalence of giardiasis in the present study was 0.42% (33/7805). Twenty-three samples (76.67%) were successfully genotyped at each locus. The bg and tpi genes had the highest typing success rate (100%). The identified assemblages were assemblage A in 27 cases (subtypes A2 and A3), and B in 3 cases.ConclusionsTo our knowledge, the present study is the first report of multilocus sequence typing of G. duodenalis isolated from humans in Romania. The present results may shed light on G. duodenalis infection in humans at a regional and national level, thus increasing awareness against this parasitic infection.

Frequency of different genotypes of Giardia duodenalis in slaughtered sheep and goat in east of iran.

Giardia duodenalis is one of the most common and important protozoan parasites of the gastrointestinal tract in humans and animals, especially in developing countries. The purpose of this study was determining prevalence of Giardia genotypes specially zoonosis genotypes in sheep and goat in eastern of iran slaughterers.This cross-sectional study was conducted during April to November 2019. 300 fecal samples were collected from the rectum of sheep and goats. The samples were subjected to DNA extraction after sucrose gradient purification. A fragment of the glutamate dehydrogenase gene (gdh) was amplified by semi-nested PCR and genotype diagnosis was performed by digestion of the secondary PCR product with restriction enzymes RsaI and Nla IV. The prevalence of Giardia was found as (274/300) by the molecular method. Restriction endonuclease digestion of the nested-PCR product showed; among 274 positive isolates, 95 were typed as assemblage E, 15 as assemblage B, 87 assemblage AI, 45 assemblage AII, and 32 assemblege C. In this study, frequency of different assemblages of G. duodenalis was determined in sheep and goats by gdh gene and PCR-RFLP method. Same of other studies, assemblage E was dominant genotype in sheep and goats. Isolation of zoonotic assemblages as AI, AII, and BIII showed that sheep and goats should be considered as a source for human infection.

Companion animals as a potential source of Giardia intestinalis infection in humans in the Czech Republic – A pilot study

Giardia intestinalis is a common enteric single-celled parasite infecting both humans and animals. Its eight morphologically identical but genetically distinct groups called assemblages differ from each other in host range. While assemblages A and B infect a wide range of hosts, including humans, the other assemblages (C to H) limit their host preferences to particular animal groups only. In companion animals as Giardia hosts, genotyping data have previously shown various results depending on pet species, location, environmental or breeding conditions, and the study design. To strengthen available epidemiological data from developed countries and to evaluate the role of pets in Giardia zoonotic transmission, we investigated Giardia-positive stool samples of three pet species (54 dogs, 18 cats, and 18 chinchillas) by a sequence-based analysis of three Giardia genes (β-giardin, glutamate dehydrogenase and triose phosphate isomerase). In dog samples, we confirmed assemblage C (21/54), assemblage D (32/54), and one case of a mixed infection C + D (1/54). In cats, we found assemblage F (16/18) and assemblage A, specifically sub-assemblage AI (2/18). All Giardia samples from chinchillas were characterised as assemblage B, specifically sub-assemblage BIV (18/18). These results indicate that in the Czech Republic, pet dogs may not represent a source of Giardia infection for humans because of the presence of only canid-specific genotypes C and D. In contrast, other pets, namely, chinchillas and, to a lesser extent, cats, may pose a potential risk of Giardia transmission to owners or breeders because they can host zoonotic Giardia genotypes.

Detection of Giardia genotypes in sewage and in stool samples in Israel

Giardia is a protozoan parasite which causes a severe disease called Giardiasis. This study was conducted to determine the prevalence of Giardia cysts in raw wastewater and its prevalence in the study community. Furthermore, the prevalence of assemblages A and B in sewage was compared with their prevalence in stool samples tested positive for Giardia. All wastewater samples were found positive for Giardia at a concentration range of 10-12225 cysts/L. Positive stool samples contained Giardia at an average concentration of 1.4x105cysts/gr. Giardia assemblage A alone was detected in 38.2% of wastewater samples, whereas assemblage B was not detected separately. However, 61.8% of the samples were found to contain both assemblages. In stool samples, the majority 27 out of 50 (54%) were found to contain assemblage B, 34.6% contained assemblage A and only 11.5% contained a mix of both assemblages. The results of the study indicate that Giardia is highly prevalent in wastewater in Israel reflecting its prevalence in the community. In addition, assemblage A is highly prevalent in wastewater, whereas assemblage B is more prevalent in stool samples, suggesting milder and/or carrier state of infection for assemblage A.

Using RFLP-PCR Technique in Determining Genotypes of Giardia lamblia from Diarrhea Cases in Children in AL-Diwaniyah City, Iraq

Introduction: Giardia lamblia is one of the most prevalent intestinal protozoa in the world, which affect children in both undeveloped and developing countries. This study aimed to determine genotypes of the Giardia lamblia using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)-PCR techniques. Additionally, the relationship between genotype patterns and their geographical distribution, gender, and age was investigated. Methods: The current study included 926 samples of faeces of children suffering from diarrhoea, who visits the internal clinics at Teaching Hospital, and Child Hospital in AL- Diwaniyah City from November 2012 - Jun 2013. For age groups of 1-12 years exclusively. The samples were examined using a direct mount wet smear, The positive samples were preserved without adding preservatives at a temperature of -20°C until the DNA extraction for G. lamblia genotyping by using PCR and RFLP-PCR technique. Results: Giardia lamblia was detected in 2.15% (20/926) of samples from diarrhea cases in children by amplification of glutamate dehydrogenase gene (gdh) using two specific primers GDHiR and GDHiF. It was revealed that 7 samples belonged to genotype A (35%) and 13 samples belonged to genotype B (65%). All genotype A samples belonged to subgenotype AII (100%), while genotype B samples belonged to subgenotypes BIII (53.61 %) and BIV (47.38 %). Genotype A was detected in children of 1-6 years of age while B genotype was detected in all age groups. Both of the genotypes have been detected in both genders (male and female) and genotype B was found in both urban and rural areas; however, its prevalence was higher in rural areas than in urban areas (100% and 30%, respectively). Conclusion: There are two genotypes of Giardia lamblia, genotype A and genotype B, each of which has secondary genetic patterns which include AII, BIII, and BIV.

Molecular Characterization of Giardia intestinalis Detected in Humans and Water Samples in Egypt

Giardia intestinalis is a common cause of gastrointestinal illness especially in children of developing countries. Giardia assemblages A and B are the major human infective genotypes. The present study aimed to investigate the role of water supply in the epidemiology of giardiasis via genotyping G. intestinalis detected in diarrheic children and in water samples in Egyptian rural areas. Stool samples of 100 diarrheic children, 40 drinking water samples and 10 raw water samples of canals were examined microscopically for Giardia. DNA was extracted from microscopically positive faecal samples and from all of the collected water samples. Amplification of Giardia tpi gene was performed by a nested PCR using assemblage A- and assemblage B-specific primers. Giardia gdh gene was amplified by a heminested PCR. Giardia genotypes were determined by restriction fragment polymorphism (RFLP) analysis of the amplified products. Sequencing of the amplified products was performed in two faecal and two water samples RESULTS: Giardia intestinalis was detected in 24 children, in none of the drinking water samples and in all canal water samples. Giardia sub-assemblage AII was identified in all stool and raw water samples. The RFLP pattern was confirmed in sequenced samples. The presence of the same Giardia sub-assemblage in diarrheic children and in raw water samples shows by molecular evidence the potential for waterborne dissemination of Giardia in Egypt. Further studies are needed to monitor cyst levels and infectivity of the genotype detected in water for risk assessment and management.

First report of Giardia duodenalis infection in the crested porcupine (Hystrix cristata L., 1758).

Italy is the only European country where the crested porcupine (Hystrix cristata) lives. A parasitological investigation was performed on faecal samples, aimed to evaluate Giardia and other parasites in a free-ranging crested porcupine population in Central Italy. Samples were collected from captured and road-killed individuals as well as from feeding areas and pathways. Collected faecal samples were examined by the Mini-FLOTAC technique and a rapid immunoassay for the search of Giardia and Cryptosporidium spp. faecal antigens. For the identification of Giardia species and genotypes, molecular analysis was performed on Giardia-positive samples, by using PCR protocols able to amplify glutamate dehydrogenase, triosephosphate isomerase and a fragment of the small subunit ribosomal RNA genes.A total of 52 crested porcupine faecal samples were collected and analysed. At microscopical examination, 39 out of 52 samples were found positive for at least a single parasite species and six different parasite taxa were identified. Forty-eight percent (25/52) of faecal samples were positive for Giardia spp. and 1.9% (1/52) for Cryptosporidium spp. at the immunoassay. Among 12 faecal samples belonging to different individuals, 33.3% (4/12) were positive for Giardia spp. By using the Mini-FLOTAC technique, positivity for Trichuris spp. (32.7%, 17/52), gastrointestinal strongyles (32.7%, 17/52), capillariid eggs (3.8%, 2/52) and coccidian oocysts (1.9%; 1/52) was also evidenced. Molecular analysis was performed on 17 out of 25 Giardia-positive isolates. At the SSU rDNA locus, expected bands were achieved for 12 out of 17 isolates and all samples were assigned to Giardia duodenalis assemblage B. Sequencing at tpi locus revealed potentially zoonotic G. duodenalis assemblage AII (two isolates) and assemblage BIV (one isolate). The present study provides the first report of G. duodenalis infection in H. cristata. More in depth studies are needed on the impact and epidemiology of G. duodenalis and other identified parasites in crested porcupines.

Prevalence and molecular characterization of Cryptosporidium spp. and Giardia intestinalis in household dogs and cats from Shiraz, Southwestern Iran.

In the present study, a total of 615 fecal samples from veterinary clinics were screened by microscopy for the presence of Cryptosporidium and Giardia oocysts. Molecular genotyping of Cryptosporidium and Giardia were carried out using PCR and sequence analysis. Overall, Cryptosporidium and Giardia oocysts were detected in the 0.6% (2/315) and 1.9% (6/315) of dogs and in the 0.7% (2/300) and 1.3% (4/300) of cats, respectively. Sequencing revealed the presence of C. canis (n = 2) in dogs and C. felis (n = 2) in cats. Moreover, G. intestinalisassemblage D (n = 2), C (n = 3) and A, sub-assemblage AII (n = 1) were identified in dogs; G. intestinalis assemblage F (n = 3) and assemblage A, sub-assemblage AI (n = 1) were identified in cats. The highest prevalence of Giardia was observed in dogs younger than one year (6/315), and in those with diarrhea (p < 0.05). Data of the study suggest that dogs and cats play a minor role in the zoonotic transmission of cryptosporidiosis and giardiasis in Southwestern Iran.