Haileeyesus Adamu

Distribution and Clinical Manifestations of Cryptosporidium Species and Subtypes in HIV/AIDS Patients in Ethiopia

Background Cryptosporidiosis is an important cause for chronic diarrhea and death in HIV/AIDS patients. Among common Cryptosporidium species in humans, C. parvum is responsible for most zoonotic infections in industrialized nations. Nevertheless, the clinical significance of C. parvum and role of zoonotic transmission in cryptosporidiosis epidemiology in developing countries remain unclear. Methodology/Principal Findings In this cross-sectional study, 520 HIV/AIDS patients were examined for Cryptosporidium presence in stool samples using genotyping and subtyping techniques. Altogether, 140 (26.9%) patients were positive for Cryptosporidium spp. by PCR-RFLP analysis of the small subunit rRNA gene, belonging to C. parvum (92 patients), C. hominis (25 patients), C. viatorum (10 patients), C. felis (5 patients), C. meleagridis (3 patients), C. canis (2 patients), C. xiaoi (2 patients), and mixture of C. parvum and C. hominis (1 patient). Sequence analyses of the 60 kDa glycoprotein gene revealed a high genetic diversity within the 82 C. parvum and 19 C. hominis specimens subtyped, including C. parvum zoonotic subtype families IIa (71) and IId (5) and anthroponotic subtype families IIc (2), IIb (1), IIe (1) and If-like (2), and C. hominis subtype families Id (13), Ie (5), and Ib (1). Overall, Cryptosporidium infection was associated with the occurrence of diarrhea and vomiting. Diarrhea was attributable mostly to C. parvum subtype family IIa and C. hominis, whereas vomiting was largely attributable to C. hominis and rare Cryptosporidium species. Calf contact was identified as a significant risk factor for infection with Cryptosporidium spp., especially C. parvum subtype family IIa. Conclusions/Significance Results of the study indicate that C. parvum is a major cause of cryptosporidiosis in HIV-positive patients and zoonotic transmission is important in cryptosporidiosis epidemiology in Ethiopia. In addition, they confirm that different Cryptosporidium species and subtypes are linked to different clinical manifestations.

Identity and public health potential of Cryptosporidium spp. in water buffalo calves in Egypt.

Little is known about the diversity and public health significance of Cryptosporidium species in water buffaloes. In this study, we examined the distribution of Cryptosporidium spp. in water buffalo calves in Egypt. Rectal fecal specimens from 179 calves and 359 adults were screened microscopically for Cryptosporidium oocysts using modified Ziehl-Neelsen stain. Cryptosporidium spp. in 17 microscopy-positive specimens from calves were genotyped by DNA sequence analysis of the small-subunit rRNA gene, and Cryptosporidium parvum was subtyped by sequence analysis of the 60 kDa glycoprotein gene. Cryptosporidium ryanae was found in 10 specimens and C. parvum in 7 specimens, with the former belonging to the newly identified C. ryanae buffalo variant and the latter belonging to the subtypes IIdA20G1 (in 5 specimens) and IIaA15G1R1 (in 2 specimens). The prevailing occurrence of C. ryanae and the subtype family IId of C. parvum and the absence of C. bovis and C. andersoni represent some features of Cryptosporidium transmission in water buffaloes in Egypt.

Prevalence of Giardia duodenalis and Cryptosporidium species infections among children and cattle in North Shewa Zone, Ethiopia

BackgroundGiardia and Cryptosporidium are the most common causes of protozoan diarrhea that lead to significant morbidity and mortality worldwide. The purpose of this study was to determine the prevalence of Giardia duodenalis and Cryptosporidium species infections among children and cattle, and to assess the potential risk of zoonotic transmission.MethodsThis cross-sectional study was conducted between January and April 2009 in Girar Jarso and Dera Districts of North Shewa Zone, Oromia Region, Ethiopia. A total of 768 stool specimens were collected and examined for intestinal parasites using direct wet mount with saline and formalin ether concentration methods. The modified Ziehl-Neelsen staining method was used for the detection of Cryptosporidium species. Statistical analysis was performed using SPSS software version 15.ResultsOut of 384 children examined, 53 (13.8%) and 28 (7.3%) were positive for Giardia and Cryptosporidium infections, respectively. Similarly, of the total 384 cattle examined, 9 (2.3%) were positive for Giardia duodenalis and 30 (7.8%) were positive for Cryptosporidium infection. The prevalence of giardiasis was significantly higher among children who had close contact with cattle 33 (18.7%) compared to children who had no contact with cattle 20 (9.6%) (P < 0.05). Higher number of Cryptosporidium infection was also recorded in children who had close contact with cattle 15 (8.5%). Difference in prevalence of giardiasis and cryptosporidiosis among children was not statistically significant between males and females. On the other hand, difference in the prevalence of giardiasis among children was statistically significant between age groups.ConclusionsHigher prevalence of Giardia duodenalis infection detected among children was significantly associated with contact with cattle and manure that the children had. Further analysis using molecular techniques is needed to explain the existence of zoonotic transmission in the study area.

Common occurrence of zoonotic pathogen Cryptosporidium meleagridis in broiler chickens and turkeys in Algeria.

Only a small number of birds have been identified by molecular techniques as having Cryptosporidium meleagridis, the third most important species for human cryptosporidiosis. In this study, using PCR-RFLP analysis of the small subunit (SSU) rRNA gene, we examined the ileum of 90 dead chickens from 23 farms and 57 dead turkeys from 16 farms in Algeria for Cryptosporidium spp. C. meleagridis-positive specimens were subtyped by sequence analysis of the 60 kDa glycoprotein gene. Cryptosporidium infection rates were 34% and 44% in chickens and turkeys, respectively, with all positive turkeys (25) and most positive chickens (26/31) having C. meleagridis. All C. meleagridis specimens belonged to a new subtype family. The frequent occurrence of C. meleagridis in chickens and turkeys illustrates the potential for zoonotic transmission of cryptosporidiosis in Algeria.

Molecular characterization of Cryptosporidium isolates from humans in Ethiopia.

In this study, 1034 faecal samples from patients with diarrhoea were screened for Cryptosporidium oocysts. Samples were collected from nine different regions in Ethiopia. Of these, 79 samples (7.6%) were positive for Cryptosporidium by modified Ziehl-Neelson staining. From all positive samples DNA was extracted and PCR amplification of the COWP, SSU-rRNA and GP60 gene fragments was performed. A total of 41 samples (52%) were positive in any of the three typing methods. The majority of isolates (39 of 41) was identified as Cryptosporidium parvum, with one Cryptosporidium hominis and one mixed infection. Sequencing of the GP60 gene fragments of 13 isolates resulted in three different subgenotypes of C. parvum, all belonging to the zoonotic subtype family IIa and one subtype of C. hominis (Ib). These data identify C. parvum as the major cause of human cryptosporidiosis in Ethiopia and suggest a zoonotic transmission of the disease in contrast to reports from other developing countries.

Prevalence and characterization of Cryptosporidium spp. in dairy cattle in Nile River delta provinces, Egypt.

Molecular characterizations of Cryptosporidium spp. in dairy cattle in industrialized nations have mostly shown a dominance of Cryptosporidium parvum, especially its IIa subtypes in pre-weaned calves. Few studies, however, have been conducted on the distribution of Cryptosporidium species and C. parvum subtypes in various age groups of dairy cattle in developing countries. In this study, we examined the prevalence and molecular characteristics of Cryptosporidium in dairy cattle in four Nile River delta provinces in Egypt. Modified Ziehl-Neelsen acid-fast microscopy was used to screen for Cryptosporidium oocysts in 1974 fecal specimens from animals of different ages on 12 farms. Positive fecal specimens were identified from all studied farms with an overall prevalence of 13.6%. By age group, the infection rates were 12.5% in pre-weaned calves, 10.4% in post-weaned calves, 22.1% in heifers, and 10.7% in adults. PCR-RFLP and DNA sequence analyses of microscopy-positive fecal specimens revealed the presence of four major Cryptosporidium species. In pre-weaned calves, C. parvum was most common (30/69 or 43.5%), but Cryptosporidium ryanae (13/69 or 18.8%), Cryptosporidium bovis (7/69 or 10.2%), and Cryptosporidium andersoni (7/69 or 10.2%) were also present at much higher frequencies seen in most industrialized nations. Mixed infections were seen in 12/69 (17.4%) of genotyped specimens. In contrast, C. andersoni was the dominant species (193/195 or 99.0%) in post-weaned calves and older animals. Subtyping of C. parvum based on sequence analysis of the 60kDa glycoprotein gene showed the presence of subtypes IIdA20G1 in nine specimens, IIaA15G1R1 in 27 specimens, and a rare subtype IIaA14G1R1r1b in one specimen. The common occurrence of non-C. parvum species and IId subtypes in pre-weaned calves is a distinct feature of cryptosporidiosis transmission in dairy cattle in Egypt. The finding of the same two dominant IIa and IId C. parvum subtypes recently found in humans in Egypt suggests calves can be potential reservoirs of zoonotic cryptosporidiosis.

High prevalence of diarrhoegenic intestinal parasite infections among non-ART HIV patients in Fitche Hospital, Ethiopia.

Background HIV infection has been modifying both the epidemiology and outcome of parasite infections. Hence, this study was undertaken to determine the prevalence of Cryptosporidium and other intestinal parasite infections among HIV positives with and without Antiretroviral Treatment(ART) and its association with CD4+ T-cell count. Methods A cross-sectional study was conducted at Fitche hospital focusing on HIV positives who came to hospital for follow-ups. A total of 378 HIV positive persons with and without ART participated in the study. Data on socio-demographic factors and diarrhoea status were obtained by interviewing all 214 with ART and 164 without ART. Stool samples were collected from all patients and examined for intestinal parasites using direct, formol-ether and modified acid-fast staining techniques. Results The prevalence of intestinal parasite infections in this study was significantly higher among HIV positive persons not on ART. Specifically, the rate of infection with Cryptosporidium species, Blastocystis spp., Giardia lamblia, and Entamoeba histolytica/E. dispar were higher, particularly in those with CD4+ T-cell counts less than 200 cells/µL. Fifty seven percent of the study participants were on ART. Out of these 164/378 (43%) of the non-ART study participants were infected with at least one intestinal parasite species. Significant association was observed between lower CD4+ T-cell count (<200 cells/µL) and the prevalence of Cryptosporidium spp. and Blastocystis spp. The two parasites were significantly more prevalent in HIV positive non-ART patients. Conclusion HIV infection increased the risk of having Cryptosporidium and other intestinal parasites and diarrhoea. Therefore, raising HIV positive’s immune status and screening for intestinal parasites is important. This study showed that patients who are taking ART had a lower prevalence of diarrhoea causing parasites and Cryptosporidium suggesting that ART through improvement of immune status of the patients may have contributed to controlling diarrhoea-causing parasites in HIV positive patients.

Prevalence and Genetic Characterization of Cryptosporidium Species in Dairy Calves in Central Ethiopia

The burden of cryptosporidiosis due to Cryptosporidium parvum is well documented in HIV-positive patients in Ethiopia. However, the role of animals in zoonotic transmission of the disease is poorly understood. The aim of this study was to determine the prevalence and genotypes of Cryptosporidium species in dairy calves; to assess the role of cattle in zoonotic transmission in central Ethiopia. A total of 449 fecal samples were collected and screened using modified Ziehl-Neelson staining method and PCR targeting the small-subunit (SSU) rRNA gene. The prevalence of Cryptosporidium was 9.4% (42/449) and 15.8% (71/449) as detected by microscopy and nested PCR, respectively. The prevalence of infection varied significantly across the study areas with the higher prevalence being observed in Chancho 25.4% (30/118). Crossbred calves had significantly higher prevalence of Cryptosporidium than indigenous zebu. Genotyping results revealed the presence of C. andersoni (76.1%), C. bovis (19.7%) and C. ryanae (4.2%). The occurrence of these Cryptosporidium species appeared to be age-related. C. andersoni constituted 92.1% of the Cryptosporidium infection in calves older than 3 months. Sequence analysis also showed the existence of intra-species variation at SSU rRNA gene. Findings of the current study indicate that cattle may not be an important source of zoonotic cryptosporidiosis in central Ethiopia. Further molecular studies are needed to support this observation from other part of the country.

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

BackgroundCryptosporidium 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.ResultsA 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, β-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, β-giardin, and glutamate dehydrogenase genes.ConclusionThe 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.

The Impact of Cooperative Social Organization on Reducing the Prevalence of Malaria and Intestinal Parasite Infections in Awramba, a Rural Community in South Gondar, Ethiopia

Introduction. Parasitic diseases are the major causes of human health problem in Ethiopia. The high prevalence of parasitic infections is closely correlated with poverty, poor environmental hygiene, and impoverished health services. Objective. The study was conducted to assess the impact of health-conscious Awramba cooperative community and its neighboring communities on the prevalence of parasitic infections in South Gondar, Ethiopia. Methods. Single stool specimens were collected from 392 individuals from Awramba and the neighboring communities. Specimens were examined microscopically for the presence of parasites using microscopy. Questionnaire was administered to determine the knowledge attitude and practice (KAP) of study participants. Results. Of the total 392 study participants examined, 58(14.8%) were positive for malaria and 173 (44.1%) for intestinal parasites. The prevalence of malaria in Awramba community (5.1%) was less than that in neighboring communities (24.5%). The prevalence of parasitic infections in Awramba (18.8%) was less than that of the neighboring communities (69.4%). Conclusion. This study showed that good household and environmental hygiene, good toilet construction and usage, and proper utilization of ITN in Awramba cooperative community have significantly contributed to the reduction of the burden of parasitic infections. Thus, the positive achievement in reducing parasitic infections in Awramba cooperative community could be used as a model for affordable health intervention in the neighboring communities, in particular, and the whole country in general.