Association of Normal and Mutated APOL1 G2 Rs60910145 alleles with SCD, Body Mass Index, and Renal Function Biomarkers and Indices.

PURPOSE OF THE STUDY: The current study aimed to detect the frequency of normal and mutated APOL1 alleles in sickle cell disease (SCD) patients and test their relation with Microalbuminuria, Creatinine, Urea, Glomerular Filtration Rate (GFR), and Body Mass Index (BMI). PATIENTS AND METHODS: The study included 156 SCD subjects. Serum Creatinine (mg/dl) and Urea (mg/dl) as well as Microalbuminuria (mg/l) level were measured by using Biosystems kit (Biosystems, Barcelona, Spain) and Mindary BA88A semi-automated biochemistry analyzer. Glomerular filtration rate and body mass index were calculated by equations. Blood DNA extraction was achieved by using the modified G-DEX™IIb Genomic DNA Extraction Kit protocol. The PCR was done for the detection of the APOL1 G2 rs60910145 alleles by using allele-specific PCR and primers. RESULTS: The CC allele was more frequent in study cases (66.7%) than TT allele. The frequency of a mutated allele (CC) was insignificantly higher in males (67.8%) than in females (65.2%) and in rural (70.9%) than urban areas. It is also higher in Shankhab compared to other tribes and subjects 26-37 years compared to other, P˃0.05. Interstingly, the subjects who carry the CC allele showed a significantly higher level of Microalbuminuria, Creatinine, BMI, and Urea compared to those carry TT allele. Moreover, GFR is also higher in subjects who carry CC than TT allele but it is not significant. CONCULSION: Altogether, the study findings highlighted the link of normal and mutated APOL1 G2 rs60910145 alleles with SCD and displayed the significant value of mutated APOL1 allele in the prediction of early nephropathy in SCD patients.

Relative risk factors for seropositive hepatitis E virus among blood donors and haemodialysis patients: The pivotal role of primary health care education.

BACKGROUND: Hepatitis E virus is a zoonotic virus with a worldwide epidemic outbreak. The aim of the study was to identify relative risk factors and co-infections concerning the seropositive HEV IgG among blood donors and haemodialysis (HD) patients in the central blood bank and renal dialysis centre in Wad Medani city, Gezira State, Sudan. MATERIALS AND METHODS: This was a cross-sectional study that included 600 participants, among them 180 showed strong seropositive HEV IgG. The structured questionnaire was used to collect data of the participants' demographics, disease risk factors and HEV IgG co-infections with HBV, HCV, HIV and syphilis. RESULTS: Among the 180 strong seropositive HEV IgG respondents, 84 were blood donors and 96 were haemodialysis patients. The gender and age (18-30 years) had a significant association with the virus exposure (P = 0.000, P = 0.000). Importantly, a significant association of HEV prevalence due to the localities effect exhibited with the highest rate among South Gezira (OR = 38, CI = 14.1-107; P = 0.000). This also observed in Wad Medani, Umm Algura, East Gezira and Managil localities (P = 0.000). The effect of the animal contact on HEV distribution exerted the significant association among the respondents for blood donors and haemodialysis patients in univariate (OR = 4.09, 95% CI 1.5-10.9; P = 0.005) and multivariate (OR = 3.2, CI = 1.1-9.4; P = 0.027) analysis. CONCLUSION: The relative risk factors of the HEV seroprevalence were gender, age, locality and animal contact. Besides the need of a regular survey for the virus seroprevalence, primary health care physicians can play pivotal role in health education, especially in rural areas of Sudan. In addition, primary health care physicians in Sudan are expected to establish strategies and plans to eradicate and minimise the health impact of HEV.

Susceptible trichostrongyloid species mask presence of benzimidazole-resistant Haemonchus contortus in cattle.

BACKGROUND: Benzimidazole (BZ) anthelmintics are widely used to control infections with parasitic nematodes, but BZ resistance is an emerging threat among several nematode species infecting humans and animals. In Sudan, BZ-resistant Haemonchus contortus populations were recently reported in goats in South Darfur State. The objective of this study was to collect data regarding the situation of BZ resistance in cattle parasitic nematodes in South Darfur using phenotypic and molecular approaches, besides providing some epidemiological data on nematodes in cattle. METHODS: The faecal egg count reduction test and the egg hatch test (EHT) were used to evaluate benzimidazole efficacy in cattle nematodes in five South Darfur study areas: Beleil, Kass, Nyala, Rehed Al-Birdi and Tulus. Genomic DNA was extracted from pools of third-stage larvae (L3) (n = 40) during trials, before and after treatment, and pools of adult male Haemonchus spp. (n = 18) from abattoirs. The polymorphisms F167Y, E198A and F200Y in isotype 1 ß-tubulin genes of H. contortus and H. placei were analysed using Sanger and pyrosequencing. RESULTS: Prevalence of gastro-intestinal helminths in cattle was 71% (313/443). Reduced albendazole faecal egg count reduction efficacy was detected in three study areas: Nyala (93.7%), Rehed Al-Birdi (89.7%) and Tulus (88.2%). In the EHT, EC50 values of these study areas ranged between 0.032 and 0.037 µg/ml thiabendazole. Genus-specific PCRs detected the genera Haemonchus, Trichostrongylus and Cooperia in L3 samples collected after albendazole treatment. Sanger sequencing followed by pyrosequencing assays did not detect elevated frequencies of known BZ resistance-associated alleles in codon F167Y, E198A and F200Y in isotype 1 ß-tubulin gene of H. placei (≤ 11.38%). However, polymorphisms were detected in H. contortus and in samples with mixed infections with H. contortus and H. placei at codon 198, including E198L (16/58), E198V (2/58) and potentially E198Stop (1/58). All pooled L3 samples post-albendazole treatment (n = 13) were identified as H. contortus with an E198L substitution at codon 198. CONCLUSIONS: To the knowledge of the authors, this is the first report of reduced albendazole efficacy in cattle in Sudan and is the first study describing an E198L substitution in phenotypically BZ-resistant nematodes collected from cattle.

The genus Madurella: Molecular identification and epidemiology in Sudan.

Eumycetoma (mycotic mycetoma) is the fungal form of mycetoma, a subcutaneous infection occurring in individuals living in endemic areas of the disease. The Sudan is hyperendemic for mycetoma, with the highest incidence being reported from Gezira State, Central Sudan. The present study was conducted at the Gezira Mycetoma Center and aimed to determine the cause of black-grain eumycetoma in the state and describe its epidemiology. Black-grain specimens were collected during the surgical operation and direct detection of the causative agent was performed using M. mycetomatis species-specific PCR and ITS PCR followed by sequencing. Black-grain was reported from 93.3% of all confirmed mycetoma cases (n = 111/119), with a prevalence in young males. Of the 91 samples subjected to direct PCR, 90.1% (n = 82) gave positive results. The predominant species (88.2%) was Madurella mycetomatis. One sample was identified as M. fahalii, one as M. tropicana, and one matched the phytopathogenic species Sphaerulina rhododendricola. The highest endemic zones were Southern Gezira (76.6%) and Northern Sinnar (23.4%). The study confirmed that direct molecular detection on grains provides rapid and specific diagnosis of agents of eumycetoma.

Vitamin D level and it is association with the severity of pulmonary tuberculosis in patients attended to Kosti Teaching Hospital, Sudan.

Globally, tuberculosis is one of the major causes of morbidity and mortality in many countries. Previous studies suggest that the incidence and severity of tuberculosis are associated with low levels of vitamin D (Vit D). Therefore, this study aimed to determine the occurrence and associated factors of vitamin D3 deficiency in pulmonary tuberculosis patients at White Nile State, Sudan. 101 individuals of diagnosed pulmonary tuberculosis patients (71 males and 30 females) and 100 non-TB controls (58 males and 42 females) were included in this study. Sputum samples were obtained from TB patients and subjected to examination for acid-fast bacilli (AFB) using Ziehl-Neelsen (ZN) stain and Gene Xpert analysis. Blood samples were collected from both groups and Serum 25(OH)-vitamin D3 was determined by an Enzyme-Linked Immunosorbent Assay. HIV infection in Tuberculosis (TB) group was also investigated using the immunochromatographic test. In our study, the majority of TB patients were suffered from TB relapse (36.6%); non-HIV infected individuals (99.1%) or showed a positive result for AFB (61.4%) in Gene Xpert analysis. Moreover, there is a significant difference in microscopy findings and bacillary levels of AFB, and Rifampicin (RIF) susceptibility pattern of M. tuberculosis strain among sputum samples of TB patients, P-values less 0.0001. Furthermore, we found that TB patients were suffered from vitamin D deficiency. In particular, the mean of vitamin D level was significantly much lower in TB patients (26.7 ± 1.6) compared to non-TB controls (117.3 ± 3.2), P-value equal 0.0001. Likewise, it's much lower in females, individuals of 21-40 years old, and patients with high bacillary levels or those infected by Rifampicin resistance strain. Accordingly, our study was highlighted the TB and Vit D deficiency relationship and showed the need for further studies to a better understanding of the impact of TB on Vit D level and investigate whether vitamin D supplementation can have a role in the prevention and treatment of tuberculosis.

New codon 198 ß-tubulin polymorphisms in highly benzimidazole resistant Haemonchus contortus from goats in three different states in Sudan.

BACKGROUND: Benzimidazole (BZ) resistance in gastrointestinal nematodes is a worldwide problem for livestock production, particularly in small ruminants. Assignment of the emergence of resistance using sensitive and reliable methods is required to adopt the correct strategies for control. In Sudan, BZ resistant Haemonchus contortus populations were recently reported in goats in South Darfur. This study aimed to provide additional data regarding albendazole efficacy and to describe the prevailing molecular BZ resistance mechanisms. METHODS: Faecal egg count reduction and egg hatch tests (EHT) were used to evaluate albendazole efficacy in three different areas of South Darfur using naturally (Rehed Al-Birdi and Tulus) and experimentally infected (Tulus and Um Dafuq) goats. Using samples from Central, East and South Darfur, pyro- and Sanger sequencing were used to detect the polymorphisms F167Y, E198A and F200Y in H. contortus isotype 1 ß-tubulin in DNA extracted from pooled third-stage larval (L3) samples (n = 36) on days 0 and 10 during trials, and from pooled adult male H. contortus (treated goats, n = 14; abattoirs, n = 83) including samples from populations previously found to be resistant in South Darfur. RESULTS: Albendazole efficacies at 5, 7.5 and 10 mg/kg doses were 73.5-90.2% on day 14 in natural and experimental infections while 12.5 mg/kg showed > 96.6% efficacy. EC50 in the EHT were 0.8 and 0.11 µg/ml thiabendazole in natural and experimental infection trials, respectively. PCRs detected Haemonchus, Trichostrongylus and Cooperia in L3 samples from albendazole-treated goats. Haemonchus contortus allele frequencies in codons 167 and 200 using pyrosequencing assays were ≤ 7.4% while codon 198 assays failed. Sanger sequencing revealed five novel polymorphisms at codon 198. Noteworthy, an E198L substitution was present in 82% of the samples (L3 and adults) including all post-treatment samples. Moreover, E198V, E198K and potentially E198I, and E198Stop were identified in a few samples. CONCLUSIONS: To our knowledge, this is the first report of E198L in BZ resistant H. contortus and the second where this is the predominant genotype associated with resistance in any strongyle species. Since this variant cannot be quantified using pyrosequencing, the results highlight important limitations in the general applicability of pyrosequencing to quantify BZ resistance genotypes.

MERS-CoV in Camels but Not Camel Handlers, Sudan, 2015 and 2017.

We tested samples collected from camels, camel workers, and other animals in Sudan and Qatar in 2015 and 2017 for evidence of Middle East respiratory syndrome coronavirus (MERS-CoV) infection. MERS-CoV antibodies were abundant in Sudan camels, but we found no evidence of MERS-CoV infection in camel workers, other livestock, or bats.

Epidemiology of strongyle nematode infections and first report of benzimidazole resistance in Haemonchus contortus in goats in South Darfur State, Sudan.

BACKGROUND: Since pastoralists in South Darfur, Sudan, had complained about lack of albendazole (ABZ) efficacy to control nematodes in goats, the frequency of infection with gastrointestinal helminths was studied before in vivo faecal egg count reduction tests (FECRT) were conducted using ABZ orally either at the dose recommended for sheep, 5 mg/kg body weight (bw) or at 10 mg/kg bw. Experiments included goats naturally infected with gastrointestinal nematodes or experimentally infected with local Haemonchus contortus isolates. Three study areas (Nyala, Beleil and Kass) were visited in autumn or winter. RESULTS: Out of 478 screened goats, 82.4% were infected with gastrointestinal helminths and 82% were shedding eggs of strongyle nematodes with 90% of the strongyle larvae representing Haemonchus spp. A FECRT using naturally infected goats (n = 225: 71 untreated, 104 and 50 treated with 5 and 10 mg ABZ/kg bw, respectively) detected reduced ABZ efficacy in Nyala and Kass. Paired and unpaired FECRT calculations detected reductions of 72-92% with samples taken at 8 days post treatment with 5 mg ABZ/kg bw and of 85-94% with 10 mg ABZ/kg bw. The FECRT based on day 14 post treatment samples showed reductions of 69-77% with 5 mg/kg and of 75-87% with 10 mg ABZ/kg bw. In Beleil, ABZ efficacy was 95%. In the egg hatch test EC50 values for Nyala and Kass ranged from 0.12-0.24 µg thiabendazole/ml, corresponding to benzimidazole resistant phenotypes. Only Haemonchus spp. larvae were present after treatments in coprocultures. When the efficacy was evaluated experimentally using isolates of H. contortus from Nyala and Kass, the 5 mg ABZ/kg dose revealed reductions of 76-78% on day 8 and of 62-70% on day 14 with the unpaired method. Using 10 mg ABZ/kg, the FECR was still only 77-82%. CONCLUSIONS: Both, in vivo and in vitro methods detected resistant H. contortus populations in goats from South Darfur State. The time point 14 days post treatment was more sensitive for detection of ABZ resistance than 8 days post treatment. This is the first report on the occurrence of anthelmintic resistance in Sudan confirming that anthelmintic resistance selection is occurring in African subsistence farming systems.

Intestinal parasitic infections among expatriate workers in various occupations in Sharjah, United Arab Emirates.

Intestinal parasitic infections are prevalent throughout many countries. This study aimed to determine the prevalence of intestinal parasite carriers among 21,347 expatriate workers, including food handlers and housemaids attending the public health center laboratory in Sharjah, UAE. Stool sample collection was performed throughout the period between January and December 2013. All samples were examined microscopically. Demographic data were also obtained and analyzed. Intestinal parasites were found in 3.3% (708/21,347) of the studied samples (single and multiple infections). Among positive samples, six hundred and eighty-three samples (96.5%) were positive for a single parasite: Giardia lamblia (257; 36.3%) and Entamoeba histolytica/Entamoeba dispar (220; 31.1%), respectively, whereas mono-infections with helminths accounted for 206 (29.1%) of the samples. Infection rates with single worms were: Ascaris lumbricoides (84; 11.9%), Hookworm (34; 4.8%), Trichuris trichiura (33; 4.7%), Taenia spp. (27; 3.81%), Strongyloides stercoralis (13; 1.8%), Hymenolepis nana (13; 1.8%), and Enterobius vermicularis (2; 0.28%), respectively. Infections were significantly associated with gender (x2 = 14.18; p = 0.002) with males as the most commonly infected with both groups of intestinal parasites (protozoa and helminths). A strong statistical association was noted correlating the parasite occurrence with certain nationalities (x2= 49.5, p <0.001). Furthermore, the study has also found a strong statistical correlation between parasite occurrence and occupation (x2= 15.60; p = 0.029). Multiple infections were not common (3.5% of the positive samples), although one individual (0.14%) had four helminth species, concurrently. These findings emphasized that food handlers with different pathogenic parasitic organisms may pose a significant health risk to the public.

A Survey of Seasonal Gastrointestinal Parasitic Infections in Donkeys from a Semiarid Sub-Saharan Region, Sudan.

Out of 92 donkeys examined for gastrointestinal parasites, 90 animals were found infected by one or more gastrointestinal parasites with an overall prevalence rate of 97.78%. The distributions of the recovered parasites in the different parts of the body were as follows: stomach, 92.4%, small intestine, 19.6%, caecum, 88%, colon, 80.4%, rectum, 73.9%, and cranial mesenteric artery, 64.1%. A significant difference was found between mean parasite counts and seasons. Hot wet season had higher mean parasites count (5411.5 ± 1694.4) in comparison with hot dry (1795.9 ± 399.6) and cool dry (1719.9 ± 522.4) seasons. Although there was no significant difference between age and mean parasite count, animals more than four years old had high mean count (3361.3 ± 921.8) in comparison with 2330 ± 744.3 and 2030.2 ± 873.1 for young and adults animals, respectively. No significant positive or negative correlation was found between total parasite counts of infected animals and any of the climatic factors. The parasites identified were Habronema spp. (40.2%), Trichostrongylus axei (30.4%), Parascaris equorum (18.5%), Anoplocephala perfoliata (4.35%), Gastrodiscus aegyptiacus (8.7%), large strongyles (84%), small strongyles (72%), and Oxyuris equi (1.1%).