Prevalence of Cryptosporidium spp. infection in a working horse population in Egypt.

Working horses support the livelihoods of smallholder farmers in Egypt. No previous study has investigated the prevalence of cryptosporidiosis in working horses in Egypt. Faecal samples were collected from 607 working horses recruited from thirty-seven villages/areas in two Egyptian governorates and examined for Cryptosporidium spp. infection using the modified Zielh-Neelsen staining technique. Data on signalment, history of recent diarrhoea, and strongyle burden were collected. The prevalence of Cryptosporidium spp. infection was calculated using a bootstrap method and potential risk factors for infection were investigated using mixed-effects logistic regression models that included sampling location as a random-effects variable. The prevalence of Cryptosporidium spp. infection was 28.7% (95% confidence interval = 23.5-33.9). None of the variables investigated, which include age, sex of the animals, and strongyle burden, were associated with risk of infection. This study provided evidence-based information on the prevalence of Cryptosporidium spp. infection in the study area. However, the potential zoonotic risk of Cryptosporidium cannot be confirmed until further studies are conducted to genotype these parasites.

Prevalence and bacterial isolation from hydatid cysts in dromedary camels (Camelus dromedarius) slaughtered at Sharkia abattoirs, Egypt.

Cystic echinococcosis (CE) is a severe neglected zoonotic parasitic disease caused by the larval stage of the dog tapeworm, Echinococcus granulosus. The objectives of this study were to determine the prevalence of hydatid cysts in dromedary camels (Camelus dromedarius) at Sharkia province, Egypt and investigate the occurrence of bacteria in hydatid fluid. A total of 6416 dromedary camels slaughtered in five abattoirs in Sharkia province, Egypt during the period from January and December 2018 were investigated for the presence of hydatid cysts. Furthermore, the bacterial species in 10 hydatid fluid isolated from lungs and livers was identified. The current findings revealed that the prevalence of hydatid cysts was 3.7%. Among those, the infection rate in lungs was 78.2%, which was significantly higher than hepatic infections (21.8%). The prevalence of hydatid cysts was the highest in winter (7.4%) and the lowest in spring (1.5%). The most common bacterial species found inside hydatid fluid collected from lungs were Salmonella spp., Staphylococcus spp., Enterococci and Pseudomonas spp. Meanwhile, Staphylococcus spp. were isolated from hepatic hydatid fluid. In conclusion, hydatid cysts infection is prevalent in dromedary camels in Sharkia province, Egypt as well as various aerobic and anaerobic bacterial species were isolated from hydatid fluid from camel lungs and livers.

Prevalence of gastrointestinal nematodes, parasite control practices and anthelmintic resistance patterns in a working horse population in Egypt.

BACKGROUND: Anthelmintic resistance is commonly reported in horse populations in developed countries, but evidence in some working horse populations is either lacking or inconclusive. OBJECTIVES: To estimate prevalence of GI nematode infections in working horses in Egypt and to evaluate strongyle resistance to ivermectin, doramectin and fenbendazole. STUDY DESIGN: Cross-sectional study. METHODS: Faecal egg count was performed on 644 working horses from 2 provinces in Egypt. A short questionnaire about horse signalment and worming history was completed for each horse. Horses identified with ≥50 strongyle type egg/g (n = 146) underwent faecal egg count reduction testing (FECRT) following treatment with ivermectin (n = 33), doramectin (n = 33) or fenbendazole (n = 30). Risk factors for strongyle (≥200 egg/g) and Parascaris equorum (>0 egg/g) infection were investigated using multivariable logistic regression analyses. RESULTS: The prevalence of low (0-199 epg), medium (200-500 epg) and high (>500 epg) strongyle infection was 88.4%, 5.9% and 5.8%, respectively. P. equorum eggs were detected in 5.1% (n = 33) of horses. Strongyle FECR was 100%, 99.97% and 100% following treatment with ivermectin, doramectin and fenbendazole respectively. Anthelmintic treatment in the 12 months preceding examination was associated with reduced likelihood of strongyle infection (odds ratio [OR] = 0.26, 95% confidence interval [CI] = 0.14, 0.47, P < .001). The likelihood of P. equorum infection was significantly associated with horses' age (OR = 0.78, 95% CI = 0.69, 0.90; P < .001). Male horses were more likely to have P. equorum infection (OR = 2.86, 95% CI = 1.37, 5.93, P = .005). MAIN LIMITATIONS: Nonrandomised selection of study areas and larval cultures was unsuccessful for some samples. CONCLUSIONS: There were low prevalence of strongyle and P. equorum infection and no evidence of macrocyclic lactones or benzimidazole resistance in strongyles in the studied working horse population.

Effect of Ginsenoside-Rh2 and Curcurbitacin-B on Cryptosporidium parvum in vitro.

Ginsenoside-Rh2 and cucurbitacin-B (CuB) are secondary metabolites of Ginseng (Panax ginseng) and Cucurbitaceae plants respectively. We assessed the anticryptosporidial activity of these two functional compounds in a cell culture model of cryptosporidiosis. The highest concentration of each compound that was not toxic to the host cells was used to assess the activity against C. parvum during infection/invasion and growth in HCT-8 cell monolayers. Monolayers were infected with pre-excysted C. parvum oocysts. Infected monolayers were incubated at 37 °C for 24 h and 48 h in the presence of different concentrations of each test compound. A growth resumption assay was performed by incubating infected monolayers in the presence of compounds for 24 h followed by a second 24-h incubation in the absence of compound. To screen for invasion inhibiting activity, freshly excysted C. parvum sporozoites were pre-treated with different concentrations of compounds prior to adding them to the cell monolayers. Paromomycin, a known inhibitor of C. parvum, and DMSO were used as positive and negative control, respectively. The level of infection was initially assessed using an immunofluorescent assay and quantified by real-time PCR. Both compounds were found to strongly inhibit C. parvum intracellular development in a dose-dependent manner. IC50 values of 25 µM for a 24 h development period and 5.52 µM after 48 h development were measured for Rh2, whereas for CuB an IC50 value of 0.169 µg/ml and 0.118 µg/ml were obtained for the same incubation periods. CuB also effectively inhibited resumption of growth, an activity that was not observed with Rh2. CuB was more effective at inhibiting excystation and/or host cell invasion, indicating that this compound also targets extracellular stages of the parasite.

Perturbation of the intestinal microbiota of mice infected with Cryptosporidium parvum.

Understanding the interaction between the intestinal microbiota (microbiome) and enteric pathogens is of interest in the development of alternative treatments that do not rely on chemotherapy and do not lead to drug resistance. We undertook research in a rodent model of cryptosporidiosis to assess whether the bacterial gut microbiota is impacted by infection with the protozoan pathogen Cryptosporidium parvum. The profile of the faecal bacterial microbiota in infected and uninfected animals was compared using 16S amplicon sequencing. In four independent experiments, the intestinal microbiota of infected mice differed from that of uninfected animals, regardless of the C. parvum isolate used to infect mice. The use of replicated treatment groups demonstrated that microbiota divergence between treatments was driven by the infection and did not result from spontaneous changes in the intestinal ecosystem unrelated to the infection. Microbiota perturbation induced by C. parvum appeared to be reversible, as we observed a tendency for the phylogenetic distance between infected and uninfected mice to diminish after mice cleared the infection. As mice infected with C. parvum do not develop diarrhoea, these observations indicate that microbiota perturbation results from other mechanisms than an accelerated movement of gut content.

Population structure of natural and propagated isolates of Cryptosporidium parvum, C. hominis and C. meleagridis.

The three protozoan species Cryptosporidium parvum, C. meleagridis and C. hominis (phylum Apicomplexa) are enteric pathogens of humans. The former two species are zoonotic and the latter is thought to infect only humans. To better characterize the structure and transmission of natural and laboratory-propagated isolates, we analyzed a collection of archived human and animal isolates of these three species by deep-sequencing polymerase chain reaction products amplified from a polymorphic sequence on chromosome 1. Thousands of screened 200-nucleotide sequences were analyzed to compare the diversity among samples, to assess the impact of laboratory propagation on population complexity and to identify taxonomically mixed isolates. Contrary to our expectation, repeated propagation in animals did not reduce intra-isolate diversity nor was diversity associated with host species. Significantly, in most samples, sequences characteristic of a different species were identified. The presence of C. hominis alleles in C. parvum and C. meleagridis isolates confirms earlier reports of mixed isolates and raises the possibility that the host range of C. hominis is broader than typically assumed. In a genetically divergent isolate of C. parvum, a majority of sequences was found to be recombinant, suggesting that this genotype originated from a C. parvum × C. hominis recombination event.