First report of knowledge and practices towards toxoplasmosis among cat owners: A cross-sectional survey in Bangladesh.

Raising awareness about Toxoplasma gondii infection among cat owners in Bangladesh is indispensable to formulate persuasive management tactics to avoid zoonotic infections from pet cats. However, to the authors' best knowledge, no studies have been performed in Bangladesh to determine knowledge and practices of toxoplasmosis in cat owners. Therefore, the objectives of the current study were to cover this research gap. We carried out a cross-sectional study in Bangladesh from June 2020 through December 2021. A structured online questionnaire was distributed to cat owners, which were voluntarily completed by them. The questionnaire included socio-demographic data, aetiology, transmissions, clinical signs, and preventive practices towards toxoplasmosis. Overall, 1,019 cat owners participated voluntarily in the cross-sectional survey. Among them, 793 (77.82%) participants showed poor knowledge regarding toxoplasmosis. Under specific knowledge sections, 62.51% of the participants revealed incorrect knowledge that toxoplasmosis was a zoonotic disease. In the same way, (72.03-85.77) % of the cat owners were unaware that the disease could be transmitted from improperly washed vegetables, raw or undercooked meat and fish, and contaminated water and milk with cat faeces. Respondents' age, education, occupation, residence type, and marital status were significantly (p < .05) associated with their knowledge level. Besides, 94.11% of cat owners had a good practice level. They followed good practices in different issues; however, they practiced those activities without knowing their impacts on disease control. Cat owners' age, education, occupation, and residence type had a significant (p < .05) association with the practice level against toxoplasmosis. This is the first study highlighting the low level of knowledge among cat owners about toxoplasmosis in Bangladesh. These knowledge gaps could increase the risk and transmission of Toxoplasma gondii infection among them and their families. The survey recommends the arrangement of educational training and programmes to increase the awareness of toxoplasmosis among cat owners.

Review on Cyclosporiasis Outbreaks and Potential Molecular Markers for Tracing Back Investigations.

Cyclosporiasis is an emerging disease caused by Cyclospora cayetanensis, which induces protracting and relapsing gastroenteritis and has been linked to huge and complicated travel- and food-related outbreaks worldwide. Cyclosporiasis has become more common in both developing and developed countries as a result of increased global travel and the globalization of the human food supply. It is not just a burden on individual human health but also a worldwide public health problem. As a pathogen of interest, the molecular biological characteristics of C. cayetanensis have advanced significantly over the last few decades. However, only one FDA-approved molecular platform has been commercially used in the investigation of cyclosporiasis outbreaks. More potential molecular markers and genotyping of C. cayetanensis in samples based on the polymorphic region of the whole genomes might differentiate between separate case clusters and would be useful in tracing back investigations, especially during cyclosporiasis outbreak investigations. Considering that there is no effective vaccine for cyclosporosis, epidemiological investigation using effective tools is crucial for controlling cyclosporiasis by source tracking. Therefore, more and more epidemiological investigative studies for human cyclosporiasis should be promoted around the world to get a deeper understanding of its characteristics as well as management. This review focuses on major cyclosporiasis outbreaks and potential molecular markers for tracing back investigations into cyclosporiasis outbreaks.

Global prevalence of Trichinella in pigs: A systematic review and meta-analysis.

BACKGROUND: Investigating the global epidemiological patterns of Trichinella in pigs is required for accurate recognition and to establishing proper control programmes and preventive measures, as well as to decrease human exposure. OBJECTIVES: To obtain a better understanding of the global prevalence of Trichinella in domestic pigs and factors that might influence the prevalence, a systematic review and meta-analysis was performed. METHODS: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed. Multiple databases were used to identify literature published between January 2000 and December 2021, representing studies from 1985 to 2021, on Trichinella prevalence in domestic pigs. Prevalence was calculated on a global and country level, by country Human Development Index (HDI), climate, pig management system, and diagnostic test. RESULTS: The global pooled prevalence based on 60 manuscripts representing 32 countries and 65 pig populations was 2.02% (95% confidence interval [CI]: 0.88-3.62) and the estimated pooled prevalence in different continents ranged from 0.00% to 11.8%. Trichinella was highest in low HDI countries (21.6%; 95% CI: 4.3-47.2), tropical wet climates (20.9%; 95% CI: 10.3-34.1), and non-intensive pig farming systems (6.1%; 95% CI: 1.0-15.5). CONCLUSIONS: While there were regional differences in Trichinella prevalence in domestic pigs, these were likely driven by country HDI and pig farming systems versus climatic factors. Increased biosecurity in outdoor pig production systems, focused meat inspections and promoting awareness could potentially decrease risk of infection.

First molecular characterization of Enterocytozoon bieneusi in children and calves in Bangladesh.

Enterocytozoon bieneusi is a widespread opportunistic pathogen found in humans and domestic animals, including cattle that poses a public health risk. This study was performed to evaluate the prevalence, genotypic diversity, and zoonotic potential of E. bieneusi among children and calves in Bangladesh. A total of 998 fecal samples were collected from children (n = 299) and calves (n = 699) and screened by nested PCR and sequencing of the ribosomal internal transcribed spacer (ITS). The overall prevalence of E. bieneusi infection was 6.4% in children and 7.9% in calves. ITS sequence analysis of 74 isolates revealed 10 genotypes, including eight known genotypes (A, D, Type IV, PigEBITS7, I, J, BEB4, and BEB6) and two new genotypes (BANEB1 and BANEB3). Specifically, genotypes A, D, Type IV, PigEBITS7, BANEB1, and BANEB3, and genotypes D, PigEBITS7, I, J, BEB4, and BEB6 were detected in children and calves, respectively. Among them, genotypes D and I were dominant genotypes in children and calves, respectively. The genotypes D and PigEBITS7 were found in both children and calves, with PigEBITS7 being observed for the first time in calves. In phylogenetic analysis, six genotypes (A, D, Type IV, PigEBITS7, BANEB1, and BANEB3), detected in 39.2% of the isolates, belonged to zoonotic Group 1. The remaining four genotypes I, J, BEB4, and BEB6 were clustered in Group 2 and are common members of the group with zoonotic potential. To the best of our knowledge, this study provides the first report of E. bieneusi infection in calves in Bangladesh and also the first molecular characterization of the parasite in children and calves in this country. Two new genotypes in children have been found, which is noteworthy. Furthermore, the presence of zoonotic genotypes indicates that cattle may serve as reservoirs for E. bieneusi, which can be a source of human microsporidiosis.

Occurrence and molecular characterization of Cryptosporidium spp. and Giardia duodenalis among captive mammals in the Bangladesh National Zoo.

Cryptosporidium and Giardia are protozoan parasites capable of causing gastrointestinal illness in humans and animals. The purpose of this research was to determine the occurrence, genetic characteristics, and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis in captive mammals at the Bangladesh National Zoo. A total of 200 fresh fecal samples from 32 mammalian species were collected and examined for Cryptosporidium spp. using nested polymerase chain reaction (PCR) targeting the small subunit (SSU) rRNA gene and G. duodenalis targeting the ß-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes. The overall infection rates of Cryptosporidium and G. duodenalis among captive mammals in the zoo were 3.5% (7/200) and 5.5% (11/200), respectively. Five species/genotypes of Cryptosporidium (C. hominis, C. andersoni, C. muris, C. felis, and Cryptosporidium deer genotype) were identified. C. hominis was subtyped as IbA12G3 by sequence analysis of the glycoprotein 60 (gp60) gene. Multilocus genotyping of G. duodenalis revealed assemblages A, B, and D. Mixed infections of assemblages B and D and A and B were found in an Asiatic jackal and a Nilgiri langur, respectively. To our knowledge, this is the first report on the occurrence and genetic identity of the two parasites among zoo animals in Bangladesh. The results suggest that zoonotic Cryptosporidium spp. and G. duodenalis are maintained in and transmitted between captive mammals. Therefore, washing, cleaning, and disinfection measures should be implemented to reduce the spread of Cryptosporidium and G. duodenalis infections.

Molecular identification and subtyping of Blastocystis sp. in hospital patients in Central China.

Blastocystis is a common enteric protist that inhabits the gastrointestinal tract of approximately 1 billion people worldwide. In this study, a total of 1,070 patients from two hospitals in Zhengzhou, Central China were enrolled to know molecular characteristics of Blastocystis sp. The microorganism was identified and subtyped with a PCR amplification and sequencing of the small subunit ribosomal deoxyribonucleic acid (SSU-rDNA). The overall minimum prevalence of Blastocystis sp. in participants was 3.1% (33/1070). Although there were no significant differences on Blastocystis sp. infections among study sites, age groups, and gender, the higher infection was observed in the patients with gastrointestinal diseases (8.8%, 15/170). Sequence analysis of the 33 isolates revealed three known subtypes, such as ST1 (n = 7), ST3 (n = 23), and ST7 (n = 3). Among them, ST3 was the dominant subtype being detected in 23 isolates (69.7%), followed by ST1 (21.2%, 7/33) and ST7 (9.1%, 3/33). The phylogenetic analysis demonstrated that three subtypes (ST1, ST3 and ST7) were clustered with their reference sequences with good bootstrap support. The subtype determination of Blastocystis sp. isolates by the phylogenetic analysis was well supported by online platform. The present study provides the first molecular report of Blastocystis sp. infections in hospital patients in Central China.

Review on parasites of wild and captive giant pandas (Ailuropoda melanoleuca): Diversity, disease and conservation impact.

The giant panda (Ailuropoda melanoleuca) is a rare species with a small global population size, and lives in the wild in only a few fragmented mountain ranges of Southwest China. Parasitic infections are among the important causes of death of giant pandas that hamper their group development. We reviewed the parasitic infections prevailing in giant pandas, and the parasitic diversity, diseases and their impact on conservation of this animal. A total of 35 parasitic species were documented in giant pandas, belonging to nematode (n = 6), trematode (n = 1), cestode (n = 2), protozoa (n = 9), and ectozoa (n = 17 (tick = 13, mite = 2, and flea = 2)). Among them, Baylisascaris schroederi had the highest prevalence and was the leading cause of death for giant pandas. Some parasites caused asymptomatic infections in giant pandas, and their health implications for the pandas remain unknown. As a whole, parasites are reported to be an important threat to the conservation of the giant pandas. Regular deworming and environmental disinfection appear to be effective ways to prevent captive giant pandas from parasitoses. In wild panda populations, parasitic control measures are suggested to include detailed examination of the ecology of the host-parasite assembly, with particular attention to density-dependent transmission. The parasitic pathogenesis and detection methods together with their biology, epidemiology, treatment, prevention and control need to be further studied for better protection of giant pandas from parasitoses.

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.

Molecular identification and subtyping of Blastocystis sp. in laboratory rats in China.

Blastocystis sp. is a ubiquitous protist that has been frequently reported in humans and animals worldwide. A total of 355 fecal samples of experimental rats were collected from four laboratory rearing facilities in China, and Blastocystis sp. was detected by PCR amplification of the partial small subunit ribosomal (SSU) rRNA gene. Twenty-nine (8.2%, 29/355) samples were positive for Blastocystis sp., with the highest infection rate (20.7%, 24/116) in rats of the Zhengzhou1, followed by that in the Zhengzhou2 (5.0%, 2/40), Shenyang (3.0%, 3/100) and Wuhan (0) rearing facilities. Among the three rat strains, Sprague-Dawley (SD) rats had higher infection rates (11.3%, 17/151) compared to Wistar rats (8.7%, 9/104) and spontaneously hypertensive (SH) rats (3.0%, 3/100). Two Blastocystis sp. subtypes (ST4 and ST7) were identified. ST4 was the predominant subtype detected in 26 samples (89.7%). A phylogenetic analysis demonstrated that the sequences of ST4 and ST7 obtained in this study were clustered with their reference subtypes. To our knowledge, this is the first report of Blastocystis sp. in experimental rats in China. Pathogen infections in laboratory animals need to be monitored due to fecal-oral transmission.

TITLE: Identification moléculaire et sous-typage de Blastocystis sp. chez des rats de laboratoire en Chine. ABSTRACT: Blastocystis sp. est un protiste omniprésent qui a été fréquemment signalé chez l'homme et les animaux du monde entier. Un total de 355 échantillons fécaux de rats de laboratoire ont été prélevés dans quatre installations d'élevage en laboratoire en Chine et Blastocystis sp. a été détectée par amplification par PCR du gène partiel de la petite sous-unité de l'ARNr ribosomique (SSU). Vingt-neuf échantillons (8,2 %, 29/355) étaient positifs pour Blastocystis sp., avec le taux d'infection le plus élevé (20,7 %, 24/116) chez les rats des élevages de Zhengzhou1, suivi de ceux de Zhengzhou2 (5,0 %, 2/40), Shenyang (3,0 %, 3/100) et Wuhan (0). Parmi les trois souches de rats, les rats Sprague-Dawley (SD) avaient un taux d'infection plus élevé (11,3 %, 17/151) que les rats Wistar (8,7 %, 9/104) et les rats spontanément hypertendus (SH) (3,0 %, 3/100). Deux sous-types de Blastocystis sp. ont été identifiés, ST4 et ST7. ST4 était le sous-type prédominant, détecté dans 26 échantillons (89,7 %). Une analyse phylogénétique a démontré que les séquences de ST4 et ST7 obtenues dans cette étude étaient groupées avec leurs sous-types de référence. À notre connaissance, il s'agit du premier signalement de Blastocystis sp. chez des rats de laboratoire en Chine. Les infections pathogènes chez les animaux de laboratoire doivent être surveillées en raison de la transmission fécale-orale.

Molecular characterization of Cryptosporidium spp. and Giardia duodenalis in experimental rats in China.

Cryptosporidium and Giardia are ubiquitous protozoan parasites that infect a broad range of hosts. The presence of Cryptosporidium spp. and G. duodenalis was detected in 355 fecal samples of laboratory experimental rats from four experimental rat rearing facilities in China by PCR amplification of the small subunit (SSU) rRNA gene. The G. duodenalis positive samples were further characterized in the ß-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes. The overall infection rates of Cryptosporidium spp. and G. duodenalis were 0.6% (2/355) and 9.3% (33/355), respectively, with no co-infection. Among the four facilities, only the rats in Zhengzhou1 were found positive for the two pathogens. Undetermined Cryptosporidium genotype was observed in one sample and C. ubiquitum in another sample. Assemblage G was identified in all the 33 G. duodenalis positive isolates at SSU rRNA gene, out of which 19, 20, and 21 isolates were also subtyped as assemblage G at tpi, gdh and bg gens, respectively. To our knowledge, this is the first report of Cryptosporidium and G. duodenalis infections in laboratory experimental rats in China. The infections of these pathogens in laboratory animals should be monitored routinely since they may interfere the biological experiments in these animals.

Evidence for Zoonotic Potential of Enterocytozoon bieneusi in Its First Molecular Characterization in Captive Mammals at Bangladesh National Zoo.

To determine the occurrence and genotypes of Enterocytozoon bieneusi in captive mammals at Bangladesh National Zoo and to assess their zoonotic significance, 200 fecal samples from 32 mammalian species were examined using a nested PCR and sequencing of internal transcribed spacer (ITS) gene. Enterocytozoon bieneusi was detected in 16.5% (33/200) of the samples. Seven different ITS genotypes were identified, including two known genotypes (D and J) and five new ones (BAN4 to BAN8). Genotype D was the most common genotype being observed in 19 isolates. In phylogenetic analysis, four genotypes (D, BAN4, BAN5, and BAN6), detected in 30 isolates (90.9%), belonged to Group 1 having zoonotic potential. The sequence of genotype J found in a Malayan pangolin was clustered in so-called ruminant-specific Group 2. The other two genotypes BAN7 and BAN8 were clustered in primate-specific Group 5. To our knowledge, this is the first report of molecular characterization of E. bieneusi in Bangladesh, particularly in captive-bred wildlife in this country. The potentially zoonotic genotypes of E. bieneusi are maintained in zoo mammals that may transmit among these animals and to the humans through environmental contamination or contact.

Molecular Characterization and Phylogenetic Analysis of Enterocytozoon bieneusi in Lambs in Oromia Special Zone, Central Ethiopia.

Enterocytozoon bieneusi is the most frequently diagnosed microsporidian species in humans and occurs in a wide range of animals. This study was conducted in Central Ethiopia to determine the prevalence and genotypes of E. bieneusi in lambs in order to evaluate their public health significance. Three hundred eighty nine fecal samples were collected and screened using a nested PCR targeting the internal transcribed spacer (ITS) of the ribosomal RNA gene. All positive PCR products were sequenced to determine the genotypes. E. bieneusi was found in 39 (10.03%) of the lambs. Differences in the infection rates among sex and age groups were not significant (P > 0.05). Five ITS genotypes belonging to three known genotypes BEB6, COS-I, and COS-II, and two novel genotypes (ET-L1 and ET-L2) were identified in lambs. All five genotypes identified in the present study clustered within cattle-specific Group 2 in the ITS phylogenetic tree. This first molecular detection and characterization of E. bieneusi in lambs in Ethiopia has identified the need for further studies in humans and other domestic animals in order to determine the public health significance of E. bieneusi in Ethiopia.

Prevalence and multilocus analysis of Giardia duodenalis in racehorses in China.

Giardia duodenalis is a zoonotic intestinal parasite infecting humans and mammals worldwide. In this study, we evaluated the prevalence of G. duodenalis in racehorses in China and genetically characterized it. In total, 621 fecal samples were collected from racehorses at 17 equestrian clubs in 15 cities in China. Forty-eight (7.7%) animals from 11 equestrian clubs were positive for G. duodenalis of assemblages A (n = 10), B (n = 36), and E (n = 2), based on the small subunit ribosomal RNA (SSU rRNA) gene. Statistically significant differences in the prevalence of this parasite were detected among the different equestrian clubs (χ2 = 49.55, df = 16, p < 0.01), whereas no significant differences were detected according to age (χ2 = 0.64, df = 1, p > 0.05) or sex (χ2 = 1.41, df = 2, p > 0.05). The G. duodenalis-positive samples were further subtyped based on three other genes, which identified 5, 4, and 4 genotypes at the triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh), and ß-giardin (bg) loci, respectively. Subassemblage BIV was the predominant genotype. A phylogenetic analysis of the concatenated sequences of subassemblage BIV showed that the multilocus genotypes from the horses were genetically different from those of humans and nonhuman primates, indicating the evolution of host separation in G. duodenalis subassemblage BIV. Our study extends our understanding of the transmission of G. duodenalis between animals and humans.

Molecular characterization of Blastocystis sp. in captive wildlife in Bangladesh National Zoo: Non-human primates with high prevalence and zoonotic significance.

Blastocystis sp. is a protozoan parasite, commonly found in the gastrointestinal tracts of animals and humans globally. The parasitic species has wide genetic diversity. Currently the mammalian and avian isolates of the parasite are grouped into 17 well known subtypes (STs), of which ten (ST1-ST9, ST12) are reported in humans. To assess the genetic diversity of Blastocystis sp. in wildlife, a total of 200 fresh fecal samples were collected from 32 mammalian wildlife species in Bangladesh National Zoo. Blastocystis sp. was screened and subtyped by PCR amplification and sequencing of the small subunit ribosomal RNA (SSU rRNA) gene. The minimum prevalence of Blastocystis sp. infection was 15.5% (31/200) in zoo animals. Eight out of 32 wildlife animal species (25.0%) were infected with Blastocystis sp. Among them, the occurrence of Blastocystis sp. was higher in non-human primates (NHPs) (31.8%) than that in herbivores (4.9%) and carnivores (0). Nucleotide sequence analysis of the SSU rRNA gene revealed seven different Blastocystis sp. subtypes, such as ST1, ST2, ST3, ST10, ST11, ST13 and ST14 in the wild animals. ST3 was the dominant subtype (41.9%, 13/31) being detected in NHPs. Of the 31 Blastocystis sp. isolates from the wild animals, 24 (77.4%) isolates belonged to the most common subtypes (ST1 to ST3) found in humans. This is the first molecular study of Blastocystis sp. in wild animals in Bangladesh. This study highlights the remarkable genetic diversity in Blastocystis sp. isolates from zoo animals and provides the first molecular evidence from spotted deer, gayal and grey langur. Due to circulation of large percentage of potentially zoonotic subtypes in the wild animals, there is a higher risk of zoonotic transmission of Blastocystis sp. in the zoo keepers and visitors.

Multilocus Typing of Enterocytozoon bieneusi in Pig Reveals the High Prevalence, Zoonotic Potential, Host Adaptation and Geographical Segregation in China.

Enterocytozoon bieneusi is one of the most frequently diagnosed Microsporidia of humans and most animals. However, there is no information on E. bieneusi infection of pigs in Tibet and Henan, China. In this study, 1,190 fecal samples were collected from pigs in Tibet and Henan and screened for the presence of E. bieneusi. The overall prevalence of E. bieneusi infection was 54.2% (645/1,190), with differences in prevalence observed among geographical areas, ages, and pig breeds. Moreover, 10 E. bieneusi genotypes were identified based on internal transcribed spacer region genotyping, including eight known genotypes (EbpC, EbpA, CHG19, CHC5, Henan-III, I, D, and H) and two novel genotypes (XZP-I and XZP-II). Multilocus sequence typing revealed 18, 7, 17, and 13 genotypes at minisatellite/microsatellite loci MS1, MS3, MS4, and MS7, respectively. Strong linkage disequilibrium (LD) and few numbers of recombination events, suggest a clonal structure of the E. bieneusi population examined in this study. The low pairwise genetic distance (FST ) and gene flow (Nm) values indicated limited gene flow in the E. bieneusi population from different hosts, with phylogenetic, structure, and median-joining network analyses all indicating the existence of host and geographical isolation. The identification of isolates belonging to nine human-pathogenic genotypes indicates that pigs play an important role in the dissemination of E. bieneusi, improving our present understanding of E. bieneusi epidemiology in the studied region.

Prevalence and genetic characterization of Cryptosporidium species and Giardia duodenalis in lambs in Oromia Special Zone, Central Ethiopia.

BACKGROUND: Cryptosporidium and Giardia duodenalis are gastro-intestinal parasites that infect human and animals worldwide. Both parasites share a broad host range and are believed to be zoonosis. The aim of this study was to identify the species of Cryptosporidium and assemblages of G. duodenalis in lambs and to elucidate their role in zoonotic transmission. RESULTS: A total of 389 fecal samples were collected from lambs and screened by microscopy and nested PCR targeting the small-subunit ribosomal RNA for Cryptosporidium; and the small-subunit ribosomal RNA, triose phosphate isomerase, ß-giardin, and glutamate dehydrogenase genes for G. duodenalis. The prevalence of Cryptosporidium and G. duodenalis was 2.1% (8/389) and 2.6% (10/389), respectively. The infection rate at the three study sites ranged from 1.3 to 3.1% for Cryptosporidium and 1.6 to 3.9% for G. duodenalis; but variation was not statistically significant (p > 0.05). The finding also showed that there is no sex and age group associated difference in the occurrence of Cryptosporidium and G. duodenalis infections in lambs. Sequence analysis revealed that lambs were mono-infection with C. ubiquitum and G. duodenalis assemblage E. The analysis also indicated the presence of genetic variation within isolates of assemblage E; with 4 of them are novel genotypes at the small-subunit ribosomal RNA, ß-giardin, and glutamate dehydrogenase genes. CONCLUSION: The findings of the current study showed that lambs are capable of harboring C. ubiquitum and G. duodenalis assemblage E. This finding suggests that lambs might be sources for potentially zoonotic Cryptosporidium species. This was first molecular study in lambs and contributes to a better understanding of the epidemiology of Cryptosporidium and G. duodenalis in central Ethiopia.

Multilocus genotyping of Giardia duodenalis isolates from calves in Oromia Special Zone, Central Ethiopia.

Giardia duodenalis is a widespread protozoan parasite that infects human and other mammals. Assessing the zoonotic transmission of the infection requires molecular characterization as there is considerable genetic variation within the species. This study was conducted to identify assemblages of Giardia duodenalis in dairy calves; and to assess the potential role of cattle isolates in zoonotic transmission in central Ethiopia. A total of 449 fecal samples were collected and screened using microscopy and PCR targeting the small-subunit (ssu) rRNA, triose phosphate isomerase (tpi), ß-giardin (bg) and glutamate dehydrogenase (gdh) genes. The overall prevalence of Giardia duodenalis in dairy calves was found to be 9.6% (43/449). The prevalence of infection based on sex, age and breed difference was statistically not significant (p>0.05). Genotyping results revealed the presence of assemblage E and assemblage A (AI). The genotypic frequency reported was 95.3% (41/43) for assemblage E and 4.7% (2/43) for assemblage A. There was one mixed infection with assemblages AI and E. Sequence analyses showed the existence of 10 genotypes within assemblage E. One genotype that showed novel nucleotide substitution was identified at the ssu rRNA locus. The other 9 genotypes, 3 at each locus, were identified at the tpi, the bg and the gdh loci with two of the gdh genotypes were novel. Findings of the current study indicate the occurrence of the livestock-specific assemblage E and the potentially zoonotic assemblage A, with the former being more prevalent. Although the zoonotic assemblage was less prevalent, there is a possibility of zoonotic human infection as AI is reported from both animals and humans.

Multilocus genotyping of Giardia duodenalis isolates from children in Oromia Special Zone, central Ethiopia.

BACKGROUND: Giardia duodenalis is the etiologic agent of giardiasis in humans and other mammals worldwide. The burden of disease is high among children in developing countries where sanitation is inadequate. However, the epidemiology and genetic diversity of this parasite is poorly understood in Ethiopia. This study aimed to determine the prevalence and genetic diversity of G. duodenalis in asymptomatic children in Oromia Special Zone, central Ethiopia. RESULTS: A total of 286 fresh fecal specimens were collected from children and screened using microscopy and PCR. The prevalence of Giardia duodenalis was 10.8 % (31/286) and 16.8 % (48/286) as detected by microscopy and nested PCR, respectively. The infection rate by the study area, sex and age group difference was not significant (P > 0.05). Genotyping results showed that 22.9 % (11/48) of the isolates belonged to assemblage A while 77.1 % (37/48) belonged to assemblage B. Although double peaks were observed at the chromatogram level, no mixed assemblage or sub-assemblage infections were demonstrated. Isolates of assemblage A mostly belonged to the sub-assemblage AII and showed similarity with previously described isolates. However, there was great genetic variability within assemblage B that showed heterogeneous nucleotide positions. Fifteen of them were new genotypes: 5 at the triose phosphate isomerase (tpi), 2 at the ß-giardin (bg), and 8 at the glutamate dehydrogenase (gdh) genes. CONCLUSIONS: Giardia duodenalis mainly assemblage B infection was predominant among the asymptomatic children in the study area. The high polymorphism found in isolates of assemblage B warrants a more defining tool to discriminate assemblage B at the sub-assemblage level. The findings of the present study indicate that there is a need to carry out national screening programs aiming to detect asymptomatic infections to minimize the reservoir of the disease.

A case report: A rare case of infant gastrointestinal canthariasis caused by larvae of Lasioderma serricorne (Fabricius, 1792) (Coleoptera: Anobiidae).

BACKGROUND: Canthariasis is a disease of humans caused by the infestation of beetle larvae. It is the second important insectal disease after myiasis. Several species of beetles are reported to cause the disease in gastrointestinal tract, urogenital system, nasal sinuses, ears and faces of mammals. The cigarette beetle Lasioderma serricorne is a widespread and destructive pest that usually feeds on tobacco, tea, beans, cereal grains, and animal and plant specimen. While there was no previous evidence of human infestation by this worm, we report the first case of L. serricorne infestation in a baby girl in China. CASE PRESENTATION: Here the case, an eight-month-old baby girl with irritable feeling, rubbing eyes, history of contact with mud and eating oranges twice during five days before attendance, and having "worms" in her stool was admitted to the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. The clinical examination revealed that the pulse rate, blood pressure and temperature were regular, and the examination of the head, neck, and chest were unremarkable. The stool specimens containing "worms" were sent to the Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University. The worms were recovered, studied morphologically using naked eyes and anatomical lens, PCR analyzed targeting cytochrome oxidase subunit 1 (COX1) and 18S rRNA genes, examined by sequence analyses of the PCR products and finally classified by phylogenetic analysis to identify their species. Based on the findings, the worms were diagnosed as the larvae of L. serricorne. CONCLUSION: This report implies that the baby had an infestation with the larvae of L. serricorne in the gastrointestine. During contact with mud or eating oranges by the girl, worm eggs were swallowed into the stomach and resisted gastric acid digestion which eventually hatched into larvae and caused canthariasis. The 8 months girl had underdeveloped immune system which might facilitate the disease. This report implicates that L. serricorne can infest human accidentally and cause canthariasis that may lead to severe damage to infant and older patient upon involvement of important organs of the body. The patients once diagnosed having canthariasis should be treated in time.

Molecular survey of Enterocytozoon bieneusi in sheep and goats in China.

BACKGROUND: Over recent years, several studies have conducted genotyping of Enterocytozoon bieneusi in various hosts worldwide using sequence analysis of the ribosomal internal transcribed spacer (ITS), however, relatively little is known about E. bieneusi in sheep and goats in China. Therefore this research was conducted to understand the prevalence and genotype distribution of E. bieneusi in farmed sheep and goats in China. RESULTS: A total of 1025 fecal specimens from farmed animals in various geographic areas were collected. Overall, PCR and sequence analysis of the ITS detected E. bieneusi in 34.4% (353/1025) of isolates; of which the prevalence in goats was 28.8% (176/611) and in sheep was 42.8% (177/414). Phylogenetic analysis of ITS sequences identified 42 genotypes (nine known and 33 novel ones). These consisted of four known genotypes (D, KIN-1, EbpC, and F) and 10 novel genotypes (CHG6, CHG7, CHG9, CHG19, CHG23, CHG25, CHS5 and CHS10-CHS12) which all belonged to the so-called zoonotic group 1. A further four known genotypes (CD6, COS-I, BEB6, and J) and 22 novel genotypes (CHG1-CHG3, CHG5, CHG8, CHG10-CHG14, CHG16-CHG18, CHG20, CHG22, CHG24, CHS3, CHS4 and CHS6-CHS9) formed a clade within the group 2. One novel genotype (CHG21) was clustered in the group 9 with the genotype CM4. CONCLUSIONS: E. bieneusi is highly prevalent, widely distributed, and genetically diverse in Chinese farmed goats and sheep. Some of the genotypes identified are potentially zoonotic.