Multiplex PCR for bacterial, viral and protozoal pathogens in persistent diarrhoea or persistent abdominal pain in Côte d'Ivoire, Mali and Nepal.

In contrast to acute diarrhoea, the aetiology of persistent digestive disorders (≥ 14 days) is poorly understood in low-resource settings and conventional diagnostic approaches lack accuracy. In this multi-country study, we compared multiplex real-time PCR for enteric bacterial, parasitic and viral pathogens in stool samples from symptomatic patients and matched asymptomatic controls in Côte d'Ivoire, Mali and Nepal. Among 1826 stool samples, the prevalence of most pathogens was highest in Mali, being up to threefold higher than in Côte d'Ivoire and up to tenfold higher than in Nepal. In all settings, the most prevalent bacteria were EAEC (13.0-39.9%) and Campylobacter spp. (3.9-35.3%). Giardia intestinalis was the predominant intestinal protozoon (2.9-20.5%), and adenovirus 40/41 was the most frequently observed viral pathogen (6.3-25.1%). Significantly different prevalences between symptomatic and asymptomatic individuals were observed for Campylobacter, EIEC and ETEC in the two African sites, and for norovirus in Nepal. Multiple species pathogen infection was common in Côte d'Ivoire and Mali, but rarely found in Nepal. We observed that molecular testing detected multiple enteric pathogens and showed low discriminatory accuracy to distinguish between symptomatic and asymptomatic individuals. Yet, multiplex PCR allowed for direct comparison between different countries and revealed considerable setting-specificity.

Microscopy detection and molecular characterisation of Giardia duodenalis infection in outpatients seeking medical care in Egypt.

Introduction: Giardiosis remains one of the most prevalent enteric parasitic infections globally. Earlier molecular-based studies conducted in Egypt have primarily focused on paediatric clinical populations and most were based on single genotyping markers. As a result, there is limited information on the frequency and genetic diversity of G. duodenalis infections in individuals of all age groups. Methods: Individual stool samples (n = 460) from outpatients seeking medical care were collected during January-December 2021 in Kafr El-Sheikh governorate, northern Egypt. Initial screening for the presence of G. duodenalis was conducted by coprological examination. Microscopy-positive samples were further confirmed by real-time PCR. A multilocus sequence typing approach targeted amplification of the glutamate dehydrogenase (gdh), beta-giardin (bg), and triose phosphate isomerase (tpi) genes was used for genotyping purposes. A standardised epidemiological questionnaire was used to gather basic sociodemographic and clinical features of the recruited patients. Results: Giardia duodenalis cysts were observed in 5.4% (25/460, 95% CI: 3.6-7.9) of the stool samples examined by conventional microscopy. The infection was more frequent in children under the age of 10 years and in individuals presenting with diarrhoea but without reaching statistical significance. Stool samples collected during the winter period were more likely to harbour G. duodenalis. All 25 microscopy-positive samples were confirmed by real-time PCR, but genotyping data was only available for 56.0% (14/25) of the isolates. Sequence analyses revealed the presence of assemblages A (78.6%, 11/14) and B (21.4%, 3/14). All assemblage A isolates were identified as sub-assemblage AII, whereas the three assemblage B sequences belonged to the sub-assemblage BIII. Patients with giardiosis presenting with diarrhoea were more frequently infected by the assemblage A of the parasite. Conclusion: This is one of the largest epidemiological studies evaluating G. duodenalis infection in individuals of all age groups in Egypt. Our molecular data suggest that G. duodenalis infections in the surveyed population are primarily of anthropic origin. However, because assemblages A and B are zoonotic, some of the infections identified can have an animal origin. Additional investigations targeting animal (domestic and free-living) and environmental (water) samples are warranted to better understand the epidemiology of giardiosis in Egypt.

Evaluación de pruebas inmunológicas en el diagnóstico de Giardia duodenalis y Cryptosporidium spp, Honduras / Evaluation of immunologic tests in the diagnosis of Giardia duodenalis and Cryptosporidium spp., Honduras

Antecedentes: No conocemos datos sobre evaluación de pruebas inmunológicas para mejorar el diagnóstico de Giardia duodenalis y Cryptosporidium spp., agentes etiológicos de diarrea de importancia mundial, en Honduras. Objetivos: Comparar dos pruebas inmunológicas para el diagnóstico de Giardia y Cryptosporidium spp. con microscopía de rutina y determinar su aplicabilidad local. Métodos: Estudio descriptivo transversal. En 2013, 134 muestras de heces recibidas en el Servicio de Parasitología del Hospital Escuela (HE) y 67 muestras del Centro de Salud Alonso Suazo (CSAS) se analizaron con una Prueba Rápida Inmunocromatográfica (PDR). En 2019-2020, 60 muestras de heces del HE se analizaron con una prueba inmunoenzimática ELISA. El protocolo de rutina incluyó examen directo en solución salina y solución de Lugol, coloración tricrómica y coloración ácido resistente modificada (ARM) (HE) y examen directo en solución salina y solución de Lugol (CSAS). Resultados: Cada prueba inmunológica mostró mayor positividad que la microscopía: en 134 muestras del HE para Giardia (6.7% vs 4.5%) y Cryptosporidium (3.7% vs 0.7%), similar en 67 muestras del CSAS (14.9% vs 7.5% para Giardia; 0.7% para Cryptosporidium con la prueba inmunológica). De 60 muestras analizadas por ELISA en HE, 31.7% fue positiva por Giardia vs 18.3% en examen directo y 23.3% en coloración tricrómica; 6.7% positiva por Cryptosporidium spp. vs 3.3% por coloración ARM. Discusión: Pruebas inmunológicas aumentaron significativamente el diagnóstico de ambas parasitosis; sin embargo, publicaciones sobre pruebas similares ofrecieron resultados no concluyentes. Por costo elevado podrían reservarse para pacientes pediátricos, pacientes inmunocomprometidos en hospitales, complementando microscopía. Los laboratorios de salud deben fortalecer capacidad diagnóstica...(AU)

Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens.

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis.Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination.Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures.Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography.Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis.Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.

Comparative analysis of routine parasitological methods for recovery of cysts, molecular detection, and genotyping of Giardia duodenalis.

In order to improve the diagnosis of giardiasis, fecal samples (high/medium/low concentration of cysts) were processed by the parasitological methods used in the routine: Faust, Lutz e Ritchie modified (replacement of formaldehyde by distilled water). The cysts were quantified; the DNA was extracted and amplified by semi-nested PCR (GDH gene). Fifteen clinical samples were analyzed to validate the study by PCR-RFLP. The results showed that the parasite was only detected and genotyped correctly when samples from children with high, medium, and low parasitic load, belonging to genotype AII, were processed by the modified Ritchie method, different from what was observed for the other methods used in laboratory routine (Faust and Lutz). The modified Ritchie method proved to be more suitable, recovering a greater number of cysts from samples, regardless of parasitic load, which reduces the chance of false negative results and has epidemiological repercussions since individuals with low parasite load are usually asymptomatic and the main disseminators of this infection.

Giardia lamblia and Helicobacter pylori coinfection in gastrointestinal biopsies: A retrospective single-center analysis from Switzerland.

BACKGROUND: The protozoan Giardia lamblia (GL) and the bacterium Helicobacter pylori (HP) are common causes of gastrointestinal disease. Coinfection is common and has been reported in studies from Africa, Europe, North America and Asia, but data for Switzerland are scarce. AIM: To investigate GL and HP prevalence and coinfection rate in gastrointestinal biopsies from the Zurich area of Switzerland. METHODS: Cases were retrieved from the laboratory information system (Medica Institute of Clinical Pathology, Zurich, Switzerland). Histological slides of cases with GL were reviewed, as were the concurrent gastric biopsies, where available. RESULTS: Between January 1, 2013 and December 31, 2020, GL was found in 88 (0.14%) of 62,402 patients with a small intestine biopsy and HP in 10,668 (15.5%) of 68,961 patients with a gastric biopsy. 74/88 (84.1%) of patients with GL had unremarkable small intestine biopsies, 13/88 (14.8%) had increased intraepithelial lymphocytes, 5/88 (5.7%) showed villous atrophy and 2/88 (2.3%) acute inflammation. 71/88 patients (80.7%) with GL had an available gastric biopsy, of which 12/71 (16.9%) were unremarkable, 28/71 (39.4%) had HP-associated gastritis, 11/71 (15.5%) showed reactive gastropathy and 1/71 (1.4%) had autoimmune gastritis. CONCLUSION: Coinfection with HP is common in patients with GL in gastrointestinal biopsies from the Zurich area of Switzerland. Therefore, gastroenterologists should consider sampling the stomach when GL is suspected for evaluation of possible concurrent HP-associated gastritis. Likewise, pathologists should scrutinize any small intestine biopsy for the presence of GL when HP-associated gastritis is seen, and vice versa.

Detection and genotyping of Giardia duodenalis from cattle and pigs in Hualien country, Eastern Taiwan.

BACKGROUND: Giardia duodenalis is a zoonotic protozoan parasite causing diarrhea through waterborne or fecal-oral infection. The cysts can live in the drinking water and cause pandemic diseases. In Taiwan, very little information is available regarding the epidemiology of G. duodenalis in domestic animals. METHODS: Fecal samples were collected from cattle (n = 156) and pigs (n = 141) in Hualien country, eastern Taiwan. Detection and genotyping were done by microscopy examination of fecal samples and amplification of the ß-giardin gene using nested PCR. RESULTS: The prevalence of G. duodenalis infection was 19.87% for cattle (31/156) and 4.26% for pigs (6/141). Using nested PCR, 30 infected samples found in cattle belonged to Assemblage E, and one sample belonged to Assemblage D. For pigs, four samples belonged to Assemblage E, one belonged to Assemblage D, and another one belonged to Assemblage A. In addition, these results showed that G. duodenalis Assemblage A was detected in pigs and may cause zoonotic transmission. CONCLUSION: This is the first epidemiological investigation of G. duodenalis infection in animals in Hualien, Taiwan. These results could provide epidemiological information for disease control and public health protection.

Occurrence of gastrointestinal parasites in dogs and cats domiciliated in Santos, SP, Brazil / Ocorrência de parasitos gastrintestinais em cães e gatos domiciliados em Santos, SP, Brasil

Abstract Helminths and protozoa are major causes of diseases in domestic animals, and many can also cause infections in humans. Knowledge of intestinal parasitoses affecting domestic animals is important for the implementation of appropriate preventive measures. The objective of this study was to evaluate the occurrence of gastrointestinal parasites in fecal samples of dogs and cats attended at the Veterinary Hospital of the Metropolitan University of Santos, SP, Brazil. We also attempted to determine whether such infection was associated with sex, age, or the presence of diarrhea. We analyzed 100 fecal samples: 85 from dogs and 15 from cats. Among the dogs, 31.8% of the samples were positive, and to 40.0% among the cats. Infection was not associated with sex or age. However, among the dogs, parasitism showed a significant association with the presence of diarrhea (P = 0.013). The helminths Ancylostoma spp. and the protozoa Giardia duodenalis were the most frequent parasites in this research. Although they present unknown species and assemblages, they are parasites with a zoonotic potential of great importance in public health. Therefore, it is essential that pets are properly diagnosed and treated against gastrointestinal parasitic infection to prevent the spread of diseases.

Resumo As enfermidades causadas por helmintos e protozoários representam uma das principais causas de doenças em animais domésticos, e muitos desses parasitos podem causar infecções em seres humanos. O conhecimento das enteroparasitoses que acometem os animais domésticos é de suma importância para que medidas preventivas adequadas sejam implementadas. O objetivo do presente trabalho foi avaliar a frequência de ocorrência de parasitos gastrointestinais em amostras de fezes de cães e gatos atendidos no Hospital Veterinário da Universidade Metropolitana de Santos, bem como sua associação com o sexo, a idade e a presença de diarreia. Do total das amostras de cães analisadas, 31,8% estavam positivas, em relação aos gatos, e 40% apresentaram positividade. Não houve associação entre o sexo e a idade, porém, foi observada associação significativa entre a presença da parasitose e da diarreia (p=0,013) entre os cães. O helminto Ancylostoma spp. e o protozoário Giardia duodenalis foram os mais frequentes na pesquisa. Embora apresentem espécies e "assemblages" desconhecidas, são parasitos de potencial zoonóticos de grande importância em saúde pública. Assim, é essencial que os animais de companhia sejam corretamente diagnosticados e tratados contra infecções parasitárias gastrintestinais para evitar a propagação de doenças.

Giardia duodenalis multi-locus genotypes in dogs with different levels of synanthropism and clinical signs.

BACKGROUND: In dogs, infections with Giardia duodenalis are mainly caused by assemblages C and D, but also by the potentially zoonotic assemblages A and B. The aims of this study were to assess differences in assemblages (i) between dogs living mainly in close proximity to humans (synanthropic dogs) versus dogs living mainly among other dogs, (ii) between samples of dogs with or without loose stool, and (iii) related to the amount of cysts shedding. METHODS: One hundred eighty-nine qPCR Giardia positive fecal samples of dogs originating from four groups (household, sheltered, hunting, and dogs for which a veterinarian sent a fecal sample to a diagnostic laboratory) were used for genotyping. For this, multi-locus genotyping of beta-giardin, triose phosphate isomerase, and glutamate dehydrogenase and genotyping of SSU rDNA gene fragments were performed. Fecal consistency was scored (loose or non-loose stool), and cysts per gram of feces were determined with qPCR. RESULTS: Assemblage D was the most prevalent in all groups, followed by the other canid assemblage C. Also, mixed C/D was common. In two (synanthropic) household dogs, the potentially zoonotic assemblage AI was present. Although occurrence of assemblage AI in household dogs was not significantly different from dogs living among other dogs (sheltered and hunting dogs), it was significantly higher compared to dogs for which a sample was sent to a diagnostic laboratory. Dogs with assemblage D shed significantly more cysts than dogs with other assemblages (except for mixed C/D results) or dogs in which no assemblage could be determined. None of the assemblages was significantly associated with loose stool. CONCLUSION: Not only do dogs mainly shed the canid Giardia duodenalis assemblages D and/or C, the numbers of cysts per gram for the canid assemblage D were also higher than for the potential zoonotic assemblage AI. Based on the assemblages shed by dogs, the risk to public health posed by dogs is estimated to be low, even though the dogs that shed AI were synanthropic household dogs. Loose stool in infected dogs was not associated with any particular Giardia assemblage.

Examination of Giardia intestinalis with Direct Microscopy and Direct Fluorescent Antibody in Patients with Diarrhea.

OBJECTIVE: In this study, our objective was to compare direct microscopic examination and direct fluorescence antibody (DFA) method for Giardia diagnosis in stool samples and to evaluate the possible risk factors related to Giardia infections. METHODS: Stool samples of 185 patients with diarrhoea collected between June 2019 and July 2019 in Erzurum Yakutiye Research Hospital were included in the study. Microscopic examination of the samples was performed with native-lugol, and they were subsequently scanned by the indirect fluorescent assay microscope using the DFA method at 100-200X magnification. In addition, all patients filled a questionnaire prepared to determine the possible risk factors related to Giardia infection. RESULTS: The age of the 185 participating patients who belonged to different groups was between 0 and 94 years. Giardia spp. cysts were detected in five stool samples (2.7%) using direct microscopic examination. Nine samples (4.9%) were DFA-positive. The incidence of giardiasis was noted to be 7.5% in children, 3.8% in adults, 7.3% in people living in rural areas, 2.9% in people living in urban areas, 10% in people having pets and 4.2% in people who do not have pets. CONCLUSION: By taking the DFA method as a reference, the sensitivity and specificity of the microscopic examination were found to be 44.4% and 99.4%, respectively. The Giardia positivity rate was higher in children, those living in rural areas, those having pets and those using well water as drinking water.

High-Yield Purification of Giardia intestinalis Cysts from Fecal Samples.

Giardia is an enteric protozoan parasite that causes gastroenteritis in all classes of vertebrates. It is ranked among the leading causes of death in children under 5 years of age. Giardiasis affects approximately 280 million people worldwide annually, a situation exacerbated by the low availability of effective treatments and the lack of a vaccine. In addition, the parasite is difficult to manipulate in in vitro environments, which hampers the development of effective disease management strategies. This article highlights the development of a method for the purification of viable Giardia cysts from fecal samples, verified by a trypan blue dye exclusion test. This protocol produces a 10-fold increase in yield over current methods. By combining sucrose flotation with gated filtration, the protocol significantly reduces the amount of debris in the purified cysts suspension. Cyst viability is verified by a trypan blue dye exclusion test. The ability to purify large quantities of Giardia from fecal samples could advance the development of effective treatments to target this worldwide prevalent parasite. © 2020 Wiley Periodicals LLC. Basic Protocol: Purification of Giardia cysts from fecal samples Support Protocol: Cyst viability test.

Giardia lamblia mimicking acute graft versus host disease after allogeneic hematopoietic stem cell transplantation: A case report.

RATIONALE: As the major complications post allogeneic hematopoietic stem cell transplantation (allo-HSCT), gastrointestinal disorders were most commonly ascribed to acute graft-versus-host disease (aGVHD) and opportunistic infections. Though Giardia lamblia (G lamblia) is the most common waterborne parasite of intestinal infection worldwide, seldom has it been reported in a patient with acute severe aplastic anemia after allo-HSCT. PATIENT CONCERNS: A 23-year-old male with severe aplastic anemia developed diarrhea, abdominal cramps, bloating, nausea, vomiting, fever, weight loss, and fatigue after allo-HSCT. DIAGNOSIS: Stool examinations for ova and parasites showed Giardia trophozoites and cysts. INTERVENTIONS: Methylprednisolone was stopped and the patient was intravenously treated with a 7-day course of metronidazole (500 mg, tid.). Simultaneously, cyclosporine (5 mg/kg) was continually utilized for suspicious gut GVHD. OUTCOMES: The Giardia lamblia in stool turned negative and his symptoms were resolved after the 7-day course. LESSONS: Incorporating non-invasive monitoring of stool examination for ova and parasites in the follow-up algorithm for post-HSCT patients can expedite clinical decision-making in the differential diagnoses for aGVHD even in the non-endemic area. Metronidazole therapy can be well-tolerated in HSCT patients with giardiasis.

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

BACKGROUND: Giardia 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. METHODS: Over 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. RESULTS: The 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. CONCLUSIONS: To 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.

Evaluation of Giardia intestinalis, Entamoeba histolytica and Cryptosporidium hominis/Cryptosporidium parvum in human stool samples by the BD MAXTM Enteric Parasite Panel.

Although the microscopic examination of stool samples remains the reference method of choice for the diagnosis of intestinal protistan infections, this method is time-consuming and requires experienced and well-trained operators. The purpose of this study was to evaluate the level of agreement between the BD MAX TM Enteric Parasite Panel (EPP) and microscopy for the detection of Giardia intestinalis (Lambl, 1859), Cryptosporidium spp. and Entamoeba histolytica Schaudinn, 1903 in stool samples. The study included faecal samples of 362 patients who were admitted to our hospital due to gastrointestinal complaints. In the microscopic examination, which was made with the native-lugol method on the stool samples that were taken from the patients, cysts, trophozoites and eggs of the parasite were examined. The diagnosis of G. intestinalis, Cryptosporidium parvum Tyzzer, 1912 and Cryptosporidium hominis Morgan-Ryan, Fall, Ward, Hijjawi, Sulaiman, Fayer, Thompson, Olson, Lal et Xiao, 2002, and E. histolytica was made in the faecal samples using the EPP assay. In the microscopic examination, Cryptosporidium spp. positive stool samples were stained with kinyoun's acid-fast. In the microscopic examination, parasites were detected in 41 (11%) of the 362 stool samples. In contrast, EPP assay identified parasites in 23 (6.3%) of the samples. In the microscopic examination, E. histolytica and Entamoeba dispar Brumpt, 1925 were detected in 22 (6.1%) of the samples, G. intestinalis was seen in 15 (4.1%), and C. parvum or C. hominis were detected in three (0.8%); these values were five (1.4%), 16 (4.4%) and two (0.5%) positive with the EPP assay. Although C. parvum or C. hominis were detected as positive in the microscopic examination of three samples, only two of the samples were positive in both EPP assay and kinyoun's acid-fast method. The EPP assay is a relatively simple test that can distinguish E. histolytica and E. dispar, but it cannot replace microscopy in the diagnosis of amoebiasis. Diagnosis for G. intestinalis and C. parvum/C. hominis with the BD MAXTM enteric parasite panel was equivalent to that with microscopy. We believe that E. histolytica must be diagnosed with nucleic acid amplification tests that have a high sensitivity and specificity like EPP assay in certain patient groups.

Detection of human intestinal protozoan parasites in vegetables and fruits: a review.

Diarrheal diseases caused by intestinal protozoan parasites are a major food-borne public health problem across the world. Vegetables and fruits provide important nutrients and minerals, but are also common sources of some food-borne human pathogenic microorganisms. The contamination of raw vegetables and fruits with human pathogenic parasites are now a global public health threat, despite the health benefits of these foods in non-pharmacological prophylaxes against diseases. A large number of reports have documented the contamination of vegetables or fruits with human pathogenic microorganisms. In this paper, we reviewed the contamination and detection methods of human pathogenic intestinal protozoans that are frequently recovered from raw vegetables and fruits. The protozoan parasites include Cryptosporidium spp., Giardia duodenalis, Cyclospora cayetanensis, Entamoeba spp., Toxoplasma gondii, Balantioides coli, Blastocystis sp., Cystoisospora belli and Enterocytozoon bieneusi. The risk factors involved in the contamination of vegetables and fruits with parasites are also assessed.

Occurrence and molecular characterization of Giardia duodenalis in lambs in Djelfa, the central steppe of Algeria.

Little is known of the prevalence and genetic identity of Giardia duodenalis in sheep in Algeria. The present study aimed at characterizing G. duodenalis in lambs up to 6 months of age in Djelfa, Algeria. A total of 346 fecal specimens were collected from 28 farms and screened for G. duodenalis cysts by zinc sulfate flotation microscopy, and positive specimens were confirmed using a direct immunofluorescence assay. Microscopy-positive specimens were analyzed by PCR and sequence analysis of the triosephosphate isomerase and glutamate dehydrogenase genes to determine G. duodenalis assemblages. Coprological examination indicated that the overall infection rate was 7.0% (24/346). Lambs under 3 months of age had higher infection rate (18/197, 9.0%) than older (6/149, 4.0%) animals, and animals with diarrhea (7/44, 16.0%) had higher infection rate than animals without diarrhea (17/302, 5.6%). PCR sequence analyses of the 15 G. duodenalis isolates revealed the presence of assemblages A in 6 isolates, assemblage E in 7 isolates, and both in 2 isolates. Assemblage A was only found in pre-weaned lambs with diarrhea, while assemblage E was mostly found in post-weaned lambs without diarrhea. The assemblage E isolates from sheep were genetically related to those from cattle in Algeria, while assemblage A isolates were from a well-known subtype prevalent in humans. Data generated from the study improve our understanding of the transmission of G. duodenalis in Algeria.

Multilocus genotyping of Giardia duodenalis from pigs in Korea.

Giardia duodenalis (syn. G. intestinalis, G. lamblia) is an important zoonotic parasite infecting livestock (including pigs) through ingesting cysts in contaminated food or water. This parasite has been classified into eight different genetic assemblages, A to H. Here, we examined the individual-level prevalence of G. duodenalis in domestic pig farms and confirmed host specificity by genotype comparisons. Samples were collected from southern and central Korea, between May 2017 and January 2019. DNA directly extracted from 745 pig fecal specimens were tested by PCR for G. duodenalis small subunit ribosomal RNA (ssu rRNA), glutamate dehydrogenase (gdh), and ß-giardin gene sequences. Based on ssu rRNA PCR, 110 (14.8%) were positive for G. duodenalis. Infection risk was the highest in the fattener group (31/139, 22.3%) and during the autumn season (52/245, 21.2%: p < .001). No statistically significant differences in risk for infection were observed between fecal types (normal versus diarrheal). Fifty ssu rRNA samples, three gdh samples, and five ß-giardin samples were successfully sequenced and genotyped. Ssu rRNA assemblage sequence analysis identified E (40.0%, 20/50), D (34.0%, 17/50), C (24.0%, 12/50), and A (2.0%, 1/50). The gdh locus identified three samples as assemblage E, and the ß-giardin locus identified four samples as assemblage E and one as assemblage C. Assemblage A sequences obtained (ssu rRNA; MK430919) had 100% identity with Giardia sequences isolated from a Korean individual (AJ293301), indicating the potential of zoonotic transmission. Continuous management and monitoring for prevention of transmission and protection of animal and human health are essential.

Intestinal parasitosis, anaemia and risk factors among pre-school children in Tigray region, northern Ethiopia.

BACKGROUND: Intestinal parasitic infections (IPIs) and anaemia are major health problems. This study assessed the prevalence of intestinal parasitic infections, anaemia and associated factors among pre-school children in rural areas of the Tigray region, northern Ethiopia. METHODS: A community based cross-sectional study was conducted among 610 pre-school children in rural communities of Northern Ethiopia from June 2017 to August 2017. Stool specimens were examined for the presence of trophozoites, cysts, oocysts, and ova using direct, formal-ethyl acetate concentration, Kato-Katz, and Ziehl-Neelsen techniques. Haemoglobin was measured using a HemoCue spectrometer. RESULTS: Among the 610 participating pre-school children in the study, the prevalence of IPIs and anaemia were 58% (95% conference interval (CI): 54.1-61.9%) and 21.6% (95% CI: 18.5-25.1%), respectively. Single, double, and triple parasitic infections were seen in 249 (41, 95% CI: 37-45%), 83 (14, 95% CI: 11-17%), and 22 (3.6, 95% CI: 2.4-5.4%) children, respectively. Of the seven intestinal parasitic organisms recorded from the participants, Entamoeba histolytica/dispar was the most prevalent 220 (36.1%) followed by Giardia lamblia 128 (20.1%), and Hymenolepis nana 102 (16.7%). Mixed infections were common among G. lamblia, E. histolytica/dispar and Cryptosporidium spp. oocyst. Intestinal parasitic infection prevalence increased from 47% in children aged 6-11 months to 66% in those aged 48-59 months; the prevalence ratio (PR) associated with a one-year increase in age was 1.08 (95% CI: 1.02-1.14, p = 0.009). Age-adjusted prevalence was higher in children who had been dewormed (PR = 1.2; 95% CI: 1.00-1.4, p = 0.045), and lower in households having two or more children aged under five (PR = 0.76, 95% CI: 0.61-0.95, p = 0.015). Anaemia rose from 28% in children aged 6-11 months to 43% in those aged 12-23 months, then fell continuously with age, reaching 7% in those aged 48-59 months. Age adjusted, anaemia was more prevalent in households using proper disposal of solid waste (PR = 1.5, 95% CI: 0.1-2.10, p = 0.009) while eating raw meat (PR = 0.49, 95% CI: 0.45-0.54, p = 0.000), any maternal education (PR = 0.64 95% CI: 0.52-0.79, p = 0.000), and household water treatment (PR = 0.75, 95% CI: 0.56-1.0, p = 0.044) were associated with lower prevalence of anaemia. CONCLUSIONS: More than half of the children were infected with intestinal parasites, while anaemia prevalence was concentrated in the 12-23 month age group. This study has identified a number of potentially modifiable risk factors to address the significant prevalence of IPIs and anaemia in these children. Improvements in sanitation, clean water, hand hygiene, maternal education could address both short and long-term consequences of these conditions in this vulnerable population.

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

BACKGROUND: 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. OBJECTIVE: 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. METHODS: 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. CONCLUSION: 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.