Epidemiology of Giardia duodenalis infection in ruminant livestock and children in the Ismailia province of Egypt: insights by genetic characterization.

BACKGROUND: Giardia duodenalis is a common flagellated protozoan parasite that infects the small intestine of a wide range of vertebrate hosts. This study aimed to determine whether tracing of G. duodenalis isolates by current genetic typing tools is possible using an exemplary set of samples from infected cattle, buffalo and children from the Ismailia province, Egypt. METHOD: A total of 804 fecal samples from ruminant animals was collected from 191 herds and 165 samples from diarrheal children below the age of 10 years. Parasites were detected in these samples using the copro-antigen RIDA®QUICK test and by real-time PCR. Samples were then genetically characterized based on the triosephosphate isomerase, glutamate dehydrogenase and ß-giardin genes. RESULTS: The prevalence of G. duodenalis was 53% in ruminants and 21% in symptomatic children and infection was not positively correlated with diarrheal symptoms. Sequence typing analysis confirmed predominance of B-type sequences (>67%) in humans and E-type sequences (>81%) in ruminants over A-type sequences. For 39 samples the complete sequence information of the three marker gene fragments could be derived. Integration of the concatenated sequence information of the three marker gene fragments with the spatial data of the respective sample revealed that identical or near identical (only up to 1 out of 1358 bp different) concatenated sequencing types were spatially related in 4 out of 5 cases. CONCLUSION: The risk of zoonotic infection emanating from ruminants even in high prevalence areas is negligible. Genetic characterization indicated a predominant anthropogenic cycle of infection within the pediatric population studied. Integration of sequence typing data with information on geographic origins of samples allows parasite sub-population tracing using current typing tools.

Comparison between two commercially available serological tests and polymerase chain reaction in the diagnosis of Cryptosporidium in animals and diarrhoeic children.

For the detection of Cryptosporidium species in 804 animals and 165 diarrhoeic children (<10 years) in Egypt, two copro-antigen tests, the RIDASCREEN® Cryptosporidium test [enzyme immunoassay (EIA)] and the RIDA®QUICK Cryptosporidium/Giardia Combi [immuno-chromatographic test (ICT)] as well as polymerase chain reaction (PCR) were used. Prevalence of Cryptosporidium was 15.0, 19.5 and 32.3% in animals and 2.4, 6.7 and 49.1% in children using EIA, ICT and PCR, respectively.Using PCR as reference method, animal samples sensitivity (Se) of the EIA was 46.5% when questionable samples were considered positive, whereas specificity (Sp) was 100%. Se of the ICT was 60.4% while Sp was 100%. Positive predictive values (PPVs) for both EIA and ICT test were 100%, and negative predictive values (NPVs) for EIA were 79.7 and 84.1% for ICT. For the children samples, the Se of EIA was 5%, Sp was 100%, PPV was 100% and NPV was 52.2%, while the Se of ICT was 13.6%, Sp was 100%, PPV was 100% and NPV was 54.6%.The Kappa score of agreement between PCR and ICT was 67.4%, 54.1% between PCR and EIA and 84.4% between ICT and EIA. Until the second serial dilution of the EIA and ICT test, 9 × 10(3) oocysts/µl of Cryptosporidia was detected, whereas in PCR, they were detected until the sixth serial dilution. Copro-antigen tests were easy to perform and less time-consuming but less sensitive compared to PCR. They obviously are best applicable for screening and epidemiological studies of large numbers of subjects, for batch specimen processing and in isolated or rural areas where reliable tests like PCR are unfeasible. When in children, a single stool sample is used for the diagnosis of clinical cases; better results can be obtained when non-standardized PCR due low specificity is coupled with copro-antigen tests.

Molecular epidemiology of Cryptosporidium in livestock animals and humans in the Ismailia province of Egypt.

The zoonotic potential of Cryptosporidium was studied in one of the most densely populated provinces of Egypt regarding livestock and people. In a representative survey, faecal samples from cattle, buffalo and stool samples from diarrhoeic children (<10 years) were investigated. Parameters assumed to be related to cryptosporidiosis were recorded for animals and children. Animal samples (804) were examined by the Copro-antigen RIDA(®)QUICK test, followed by PCRs targeting the 18S rDNA and gp60 genes for antigen-positive and 10% randomly selected negative samples. All 165 human samples were tested by both methods. The overall estimated prevalence of Cryptosporidium in ruminants was 32.2%, without significant difference between animal species. PCR identified 65.7% Cryptosporidium parvum, 11.8% Cryptosporidium ryanae, 4.1% Cryptosporidium bovis, and combinations of C. parvum plus C. ryanae (11.2%), C. parvum plus C. bovis (5.3%) and of C. parvum plus Cryptosporidium andersoni (1.8%), also without significant differences in species occurrence between cattle and buffalos. The human Cryptosporidium spp. prevalence was 49.1%, of which 60.5% were Cryptosporidium hominis, 38.2% C. parvum and 1.2% C. parvum plus C. bovis. Analysis of gp60 variants allocated C. parvum found in animals to the zoonotic subtype family IIa (18.9%, subtype IIaA15G1R1 only) and to IId (81.1%, mostly IIdA20G1). In humans 50% were classified as subtype family IIa (IIaA15G1R1 and IIaA15G2R1) and 50% were IIdA20G1. C. andersoni occurred only in cattle older than 1 year. In contrast, mono-infections with one of the three single Cryptosporidium species and the three combinations with C. parvum were more prevalent in cattle and buffaloes younger than 1 year, particularly in those younger than 3 months, and were predominantly subtype family IId. In human samples no Cryptosporidium were identified in children younger than 7 months. Neither place of residence nor the source of drinking-water had measurable effects on prevalence. Remarkably, however, all children with C. parvum subtype family IIa and 86% with subtype family IId had contact to animals. High prevalence and identical genotypes of C. parvum in animals and humans indicate zoonotic transmission due to contact with animals, involving IIdA20G1 as the most frequent subtype.

Prevalence of Salmonella in retail chicken meat in Hanoi, Vietnam.

Infection with Salmonella (S.) is the most frequently reported cause of bacterial food-borne illness worldwide. Poultry are a common source and, in recent years, much attention has been focused in determining the prevalence of Salmonella during the different stages in the poultry production chain. This article was designed to investigate the prevalence of Salmonella serovars in retail chicken meat sold in Hanoi. A total of 262 samples were randomly collected from retail markets and examined for the presence of Salmonella. Of these samples, 48.9% were found to be contaminated with Salmonella. Predominant serotypes were S. Agona, S. Emek, S. London. The prevalence of S. Enteritidis and S. Typhimurium was considered. These findings have highlighted the magnitude of Salmonella contamination in retail chicken meat in Hanoi. On the basis of these preliminary survey results, it is recommended that a cost-effective monitoring and surveillance system for Salmonella should be established in Hanoi. This system should be augmented by good agricultural and hygienic practices and well-designed longitudinal research activities on the whole poultry production chain.

Preliminary analysis of tetracycline residues in marketed pork in Hanoi, Vietnam.

A cross-sectional survey was designed to investigate the proportion of tetracycline residues in marketed pork in suburb and urban districts in Hanoi. A total of 290 raw muscle samples were randomly collected from open markets in these districts. The samples were qualitatively screened for tetracycline residues using the agar inhibition test, and Bacillus cereus (ATCC 11778) as the reference strain. The inconclusive samples were then analyzed using high-performance liquid chromatography (HPLC). The positive samples from either test were defined as positive results. Overall, 5.5% of all collected samples were positive for tetracycline residues. The proportion of positive samples from shops in suburb districts was significantly (P < 0.05) different from those collected from shops in urban districts. So, the factor of region was identified as a risk factor of tetracycline residue proportion in raw pork with an odds ratio (OR) of 4.03 (95% CI = 1.12, 14.45). For the other factors, such as season, type of shop, type of abattoir, origin of meat, etc., the difference in proportion of positive samples within each factor was substantial but not statistically significant. These factors were identified as nonrisk factors. Such a high proportion may pose a potential hazard to public health, particularly since they might induce drug resistance of pathogenic micro-organisms.

The phytochemical profile and identification of main phenolic compounds from the leaf exudate of Aloe secundiflora by high-performance liquid chromatography-mass spectroscopy.

The phytochemical profile of Aloe secundiflora (Aloeaceae) and the identity of eight major compounds, including the two main constituents, have been determined from the leaf exudate of this ethnoveterinary used species from Kenya and Tanzania. Analytical HPLC-MS studies of the exudate have revealed that it comprises a mixture of phenolic compounds, mainly anthrones (aloenin, aloenin B, isobarbaloin, barbaloin and other aloin derivatives), chromones and phenylpyrones with a low content of polysaccharides and aliphatic compounds. The high percentage of anthrones in the exudate could provide a first line of evidence for the use of the plant in ethnoveterinary practices.