Severe coinfection of dengue and malaria: A case report.

Key Clinical Message: In countries like Sudan, where several infectious diseases are prevalent, health care providers should not be satisfied with initial detection of a single pathogen and whenever it is feasible, they should investigate coinfections. Infections with high mortality or severe morbidity should be prioritized during the differential diagnosis particularly for diseases with similar clinical manifestations to reduce the death and disability rates. However, this requires substantial improvement in the diagnostic capacity. Abstract: Here we report a case of dengue and malaria coinfection from the southeast region of Sudan, bordering Ethiopia and Eritrea. A 25-year-old male from Sudan presented with symptoms of fever, chills, vomiting, and muscle and joint pain. Laboratory investigations confirmed a coinfection of dengue and malaria, which is assumingly not uncommon in areas heavily syndemic with several diseases but it is severely under-detected, underreported, and underestimated. The case has fully recovered after the supportive care for dengue and chemotherapy treatment for malaria. In such a case, it was important to monitor the patient's recovery and the treatment outcome through clinical indicators and laboratory parameters to update the treatment course whenever needed, according to response. The increasing burden and outbreaks of vector-borne diseases including dengue and malaria in Sudan, indicates the need for improving the implementation of the global vector control response that established by the World Health Organization. Additionally, the increasing prevalent of coinfections is urging substantial improvement in the diagnostic capacity in endemic countries.

Intra- and interspecies variation and population dynamics of Fasciola gigantica among ruminants in Sudan.

Fasciola gigantica is a widespread parasite that causes neglected disease in livestock worldwide. Its high transmissibility and dispersion are attributed to its ability to infect intermediate snail hosts and adapt to various mammalian definitive hosts. This study investigated the variation and population dynamics of F. gigantica in cattle, sheep, and goats from three states in Sudan. Mitochondrial cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 1 (ND1) genes were sequenced successfully to examine intra and interspecific differences. ND1 exhibited higher diversity than COI, with 15 haplotypes and 10 haplotypes, respectively. Both genes had high haplotype diversity but low nucleotide diversity, with 21 and 11 polymorphic sites for ND1 and COI, respectively. Mismatch distribution analysis and neutrality tests revealed that F. gigantica from different host species was in a state of population expansion. Maximum likelihood phylogenetic trees and median networks revealed that F. gigantica in Sudan and other African countries had host-specific and country-specific lineages for both genes. The study also indicated that F. gigantica-infected small ruminants were evolutionarily distant, suggesting deep and historical interspecies adaptation.

Delayed cerebellar ataxia induced by Plasmodium falciparum malaria: A rare complication.

Key Clinical Message: In endemic areas, malaria-induced cerebellar ataxia should be suspected in patients presenting with neurological disorders including slurred speech, tremors, and a sense of imbalance and dizziness while walking. Healthcare providers should be aware to properly investigate and early detect and manage infections associated with the development of cerebellar ataxia to improve the case management and clinical outcome cost-effectively. Abstract: Here, we report the clinical manifestations, investigations, and outcomes of a patient developed delayed cerebellar ataxia following a malaria infection: an unusual complication of the disease. This report highlights the diagnostic challenges in a country endemic with several infectious diseases, yet it has a limited diagnostic and surveillance capacity.

Molecular detection of Epstein-Barr virus among Sudanese patients diagnosed with Hashimoto's thyroiditis.

OBJECTIVES: Hashimoto's thyroiditis (HT) is the most common cause of hypothyroidism. The exact mechanism initiating the development of HT is not yet clear. This study aimed to investigate the correlation between HT and the presence of Epstein-Barr virus (EBV) in a Sudanese population. RESULTS: EBV-LMP1 was detected in 11.1% of HT cases, which is consistent with previous studies. Studies have reported a wide range of frequencies indicating the presence of EBV in HT, and patients with autoimmune thyroiditis have increased titers of anti-EBV antibodies in their sera compared to healthy subjects. Intrathyroidal EBV-infected B cells may be responsible for the increased risk of development of B-cell lymphoma in the thyroid gland in patients with autoimmune thyroiditis. Our study suggests that regular follow-up is necessary for patients diagnosed with HT and are positive for EBV, as antiviral therapy is not applicable due to the risk of thyroid dysfunction. The study suggests an association between EBV and HT, but causation cannot be determined. The study also highlights the need for further research to determine the viral role and correlate it with the severity and progression of HT.

Rift Valley fever and malaria co-infection: A case report.

Key Clinical Message: We report a case of febrile illness that was presented with mild symptoms. However, laboratory investigation confirmed a malaria and Rift Valley fever co-infection. Healthcare providers in settings endemic with several infectious diseases should seek rolling out possibilities of other infections prior to starting treatment for achieving effective case management with less resources and better safety of patients. Abstract: Here we report a case of febrile illness that confirmed to be a co-infection of malaria and Rift Valley fever. The patient was initially diagnosed with malaria and started on treatment immediately. However, due to the lack of response to the treatment further laboratory investigations were pursued.

Detection of Nonsynonymous Single Variants in Human HLA-DRB1 Exon 2 Associated with Renal Transplant Rejection.

Background: HLA-DRB1 is the most polymorphic gene in the human leukocyte antigen (HLA) class II, and exon 2 is critical because it encodes antigen-binding sites. This study aimed to detect functional or marker genetic variants of HLA-DRB1 exon 2 in renal transplant recipients (acceptance and rejection) using Sanger sequencing. Methods: This hospital-based case-control study collected samples from two hospitals over seven months. The 60 participants were equally divided into three groups: rejection, acceptance, and control. The target regions were amplified and sequenced by PCR and Sanger sequencing. Several bioinformatics tools have been used to assess the impact of non-synonymous single-nucleotide variants (nsSNVs) on protein function and structure. The sequences data that support the findings of this study with accession numbers (OQ747803-OQ747862) are available in National Center for Biotechnology Information (GenBank database). Results: Seven SNVs were identified, two of which were novel (chr6(GRCh38.p12): 32584356C>A (K41N) and 32584113C>A (R122R)). Three of the seven SNVs were non-synonymous and found in the rejection group (chr6(GRCh38.p12): 32584356C>A (K41N), 32584304A>G (Y59H), and 32584152T>A (R109S)). The nsSNVs had varying effects on protein function, structure, and physicochemical parameters and could play a role in renal transplant rejection. The chr6(GRCh38.p12):32584152T>A variant showed the greatest impact. This is because of its conserved nature, main domain location, and pathogenic effects on protein structure, function, and stability. Finally, no significant markers were identified in the acceptance samples. Conclusion: Pathogenic variants can affect intramolecular/intermolecular interactions of amino acid residues, protein function/structure, and disease risk. HLA typing based on functional SNVs could be a comprehensive, accurate, and low-cost method for covering all HLA genes while shedding light on previously unknown causes in many graft rejection cases.

Knowledge and attitudes towards cervical cancer prevention among women in Khartoum state, Sudan.

BACKGROUND: Cervical cancer is considered the third leading cause of death among women worldwide, and human papillomavirus was identified as a major causative agent for developing cervical cancer. OBJECTIVES: This study aimed to assess the knowledge and attitudes towards cervical cancer prevention among women in Khartoum state, Sudan. DESIGN: A community-based cross-sectional study implemented in Khartoum state, Sudan, from 1 August 2020 to 1 September 2020. METHODS: We conducted a descriptive cross-sectional community-based study using an electronic questionnaire for data collection. Descriptive statistics, frequency, mean, and percentage were computed. RESULTS: The study included 716 female participants with a mean age of 27.6 + 8.7 years. 580 (81.0%) and 229 (32.0%) had heard about cervical cancer and Pap test, respectively. cervical cancer was assumed related to alcohol consumption 109 (15.2%), giving birth to many children 51 (7.1%), ageing 118 (16.5%), and having many sexual partners 335 (46.8%). In addition, 300 (41.9%) attributed cervical cancer to having human papillomavirus infection, 256 (35.6%) to the prolonged use of contraceptives, and 162 (22.6%) to smoking. Knowledge about the best time to be vaccinated against human papillomavirus, 110 (15.4%) stated it is better after marriage. Regression models to predict the effectors on participants' knowledge and attitudes showed a low standard deviation of the estimates with higher values of the adjusted R2 [R: 0.041, 0.017, and 0.006; std: 1.527, 0.417, and 0.426]. This indicates the combined influence of occupation, educational level, family income, and marital status on the participant's knowledge and attitude levels. CONCLUSION: This study revealed that the participant's knowledge and attitudes levels are mainly driven by their occupation, educational level, family income, and marital status altogether. This underscores the need for a countrywide community engagement campaign through health education and raising awareness sessions, and massive social media to sensitize the community and healthcare providers about the risk of cervical cancer and the available prevention and control measures.

Genetic variation and demographic history of Sudan desert sheep reveal two diversified lineages.

More than 400 million sheep are raised on the African continent, the majority of which are indigenous and are primarily reared for sustenance. They have effectively adapted to various climatic and production environments, surviving and flourishing. The genetic relationships among these sheep populations remain understudied. Herein, we sequenced the entire mitochondrial DNA control region of 120 animals from Hamary and Kabashi and their crossbreed (Hamary x Kabashi) of Sudan desert sheep (SDS) to understand their maternal-inherited genetic variation and demographic history profiles and relate those to the history of sheep pastoralism on the African continent. The results show a diversified and predominant D- loop haplogroup B (n = 102, 85%), with all other sequences belonging to haplogroup A. Most of the maternal genetic variation was partitioned between haplogroup (76.3%) while within haplogroup accounted for 23.7% of the variation. However, little genetic differentiation was observed among the two breeds and their crosses, with our results supporting a Hamari maternal origin for the crossbreed. Bayesian coalescent-based analysis reveals distinct demographic history between the two haplogroups, two breeds and their crosses. Comparison of the two haplogroup showed that haplogroup B experienced an earlier expansion than haplogroup A. Unlike the breed-based comparison, the expansion of the two breeds started roughly at the same time, around 6500 years ago, with Kabashi having a slightly greater effective population size. The maternal ancestors of SDS may have diverged before their introduction to the African continent. This study provides novel insights into the early history of these two main breeds of Sudan desert sheep and their crosses.

Guillain-Barre syndrome associated with COVID-19 infection: A case series.

In this communication, we reported a series of six patients presented with Guillain-Barré syndrome that associated with COVID-19 infection, which was confirmed with RT-PCR. Here we discuss the laboratory investigation and case management, as well as clinical presentation and outcome of each case. The current report demonstrated the first case series of COVID-19-associated GBS-cases in Sudan.

The First Molecular Detection of Aedes albopictus in Sudan Associates with Increased Outbreaks of Chikungunya and Dengue.

As part of our surveys of the invasive malaria vector Anopheles stephensi in four Sudanese states, including North and South Kordofan, Sennar, and White Nile, we collected 166 larvae. Our morphological identification confirmed that 30% of the collected mosquito samples were Anopheles species, namely An. gambiae s.l. and An. stephensi, while the 117 Aedes specimens were Ae. luteocephalus (39%), Ae. aegypti (32%), Ae. vexans (9%), Ae. vittatus (9%), Ae. africanus (6%), Ae. metalicus (3%), and Ae. albopictus (3%). Considering the serious threat of Ae. albopictus emergence for the public health in the area and our limited resources, we prioritized Ae. albopictus samples for further genomic analysis. We extracted the DNA from the three specimens and subsequently sequenced the cytochrome oxidase 1 (CO1) gene and confirmed their identity as Aedes albopictus and their potential origin by phylogenetic and haplotype analyses. Aedes albopictus, originating from Southeast Asia, is an invasive key vector of chikungunya and dengue. This is the first report and molecular characterization of Ae. albopictus from Sudan. Our sequences cluster with populations from the Central African Republic and La Réunion. Worryingly, this finding associates with a major increase in chikungunya and dengue outbreaks in rural areas of the study region and might be linked to the mosquito's spread across the region. The emergence of Ae. albopictus in Sudan is of serious public health concern and urges for the improvement of the vector surveillance and control system through the implementation of an integrated molecular xenosurveillance. The threat of major arboviral diseases in the region underlines the need for the institutionalization of the One Health strategy for the prevention and control of future pandemics.

Gene polymorphisms of cyclin-dependent kinase inhibitor and matrix metalloproteinase-9 in Sudanese patients with esophageal squamous cell carcinoma.

BACKGROUND: The polymorphisms of the cyclin-dependent kinase inhibitor (CDKN1A) gene and matrix metalloproteinase-9 (MMP9) gene may increase one's susceptibility to malignancies. In this study, the association of the single nucleotide polymorphisms (SNPs) CDKN1A rs1059234 c.70C>T at the 3' untranslated region and MMP9 rs17576 (c.836A>G, p.Gln279Arg) with esophageal squamous cell carcinoma (ESCC) in Sudanese individuals were investigated. MATERIALS AND METHODS: A case-control study involving age- and gender-matched groups were conducted in a cancer center in eastern Sudan (Gadarif) between April and October 2020. The case group consisted of ESCC patients, whereas the control group comprised healthy subjects. Polymerase chain reaction-restriction fragment length polymorphism was performed for the genotyping of the CDKN1A rs1059234 and MMP9 rs17576 SNPs. The genotyping results were confirmed by Sanger sequencing. RESULTS: The genotype distributions for CDKN1A rs1059234 and MMP9 rs17576 were in agreement with the Hardy-Weinberg equilibrium. The variant allele T in CDKN1 rs1059234 c.70C>T was significantly more prevalent in the ESCC patients than in the healthy controls [51.3% vs. 19.2%; OR = 4.4; 95% CI (2.6-7.4); p < 0.001]. Moreover, in CDKN1A rs1059234, the genotype TC + TT [76.9% vs. 38.4%; OR = 5.3; 95% CI (2.6-10.7); p < 0.001] was more frequent in the cases than in the controls, and it was significantly associated with ESCC risk. In MMP9 rs17576, the variant allele G was also significantly prevalent in the cases relative to the controls, and it was significantly associated with increased ESCC risk in the cases compared with the controls [27.5% vs. 1.9%; OR = 19.4; 95%CI (5.8-64.1); p < 0.001]. Both genotypes containing the allele G (AG + GG) were the most common genotypes in the cases [48.7% vs. 3.8%; OR = 23.7; 95%CI (6.8-81.7); p < 0.001], and they significantly increased the risk of ESCC. CONCLUSION: A significant increase in ESCC risk is associated with the SNPs CDKN1A rs1059234 and MMP9 rs17576.

Hepatitis E Virus Outbreak among Tigray War Refugees from Ethiopia, Sudan.

We report hepatitis E virus (HEV) outbreaks among refugees from Ethiopia in Sudan during June 2021-February 2022. We identified 1,589 cases of acute jaundice syndrome and used PCR to confirm HEV infection in 64% of cases. Implementing vaccination, water, sanitation, and hygiene programs might reduce HEV outbreak risk.

The use of cholera oral vaccine for containment of the 2019 disease outbreak in Sudan.

A cholera outbreak in Blue Nile and Sennar states, south-eastern and southern Sudan, took place during September-December 2019. An outbreak surveillance sample collection was made. Vibrio cholerae O1 Ogawa was isolated from clinical samples of all confirmed 200 and 132 cases in Blue Nile state and Sennar state, respectively. The case fatality rate was higher in Blue Nile state, 4% compared with only 2.3% in Sennar state. The Euvichol-Plus oral cholera vaccine was rapidly deployed for the first time in Sudan to the most at-risk populations in the two affected states, 1 471 188 and 1 546 542 individuals in Sennar and Blue Nile states, respectively. The rapid deployment of cholera vaccines as the major prevention and control strategy was successful and helped greatly with the containment of this epidemic. In-depth genomics studies are crucial for understanding the disease dynamics in Sudan by identifying locally circulating strains of the bacteria and further improving prevention and control strategy by characterising the susceptibility and resistance of these locally circulating strains to currently used antibiotics.

Blood meal profile and positivity rate with malaria parasites among different malaria vectors in Sudan.

BACKGROUND: Malaria is a life-threatening public health problem globally with particularly heavy burden in the sub-Saharan Africa including Sudan. The understanding of feeding preference of malaria vectors on different hosts is a major challenge for hindering the transmission cycle of malaria. In this study, blood meals taken by blood-fed Anopheles mosquitoes collected from the field in malaria endemic areas of Sudan were analysed for source of blood meal and malaria parasite presence. METHODS: Anopheles mosquitoes were collected from different regions in Sudan: Khartoum state, Sennar state, Northern state, and El Gedarif state between September 2020 and February 2021. Anopheles mosquitoes were collected using the standard pyrethrum spray catch and back-pack aspirator. Mosquito samples were sorted and morphologically identified to species level using international identification keys. Morphologically identified mosquito species were also confirmed using PCR. Genomic DNA was extracted from mosquitoes for molecular identification of blood meal source and parasite detection. The presence of Plasmodium species DNA in each mosquito sample was investigated using semi-nested PCR. Frequency of each blood meal source, Anopheles mosquito vector, and malaria parasite detected was calculated. Positivity rate of each fed female Anopheles mosquito was calculated for each species. RESULTS: A total of 2132 Anopheles mosquitoes were collected. 571 (26.8%) were males and 1561 (73.2%) were females classified based on their abdominal status into 1048 (67.1%) gravid, 274 (17.6%) fed, and 239 (15.3%) unfed females. Among the blood fed Anopheles mosquitoes, 263 (96.0%) were morphologically identified and confirmed using PCR to Anopheles arabiensis, 9 (3.3%) to Anopheles stephensi, and 2 (0.7%) to Anopheles rufipes. Of 274 blood-fed An. arabiensis, 68 (25.9%) fed on mixed blood meals from human and cattle, 8 (3.0%) fed on cattle and goat, and 13 (4.8%) fed on human, cattle and goat. For single blood meal sources, 70 (26.6%) fed on human, 95 (36.1%) fed on cattle, 8 (3.0%) fed on goat, and 1 (0.4%) fed on dog. While An. rufipes and An. stephensi fed on dog (2; 0.75%) and cattle (9; 3.3%), respectively. Plasmodium parasite detection in the blood meals showed that 25/274 (9.1%) An. arabiensis meals were positive for Plasmodium vivax and 19/274 (6.9%) An. arabiensis meals were positive for Plasmodium falciparum. The rate of positivity of An. arabiensis with any Plasmodium species was 16.7%. However, the positivity rate with P. falciparum only was 7.2%, while P. vivax was 9.5%. Both An. rufipes and An. stephensi were having positivity rates of 0.0% each. CONCLUSIONS: This study which was mainly on blood-fed Anopheles mosquitoes showed a diversity in the type of diet from human, cattle, and goat. Anopheles mosquitoes especially An. arabiensis in Sudan, are opportunistic blood feeders and can feed broadly on both human and cattle. The application of blood meal identification is not only important in malaria vector epidemiological surveillance but also is very useful in areas where arthropods exhibit zoophilic feeding behaviour for mammals.

The Phylodynamic and Spread of the Invasive Asian Malaria Vectors, Anopheles stephensi, in Sudan.

Anopheles stephensi is an invasive Asian malaria vector that initially emerged in Africa in 2012 and was reported in Sudan in 2019. We investigated the distribution and population structure of An. stephensi throughout Sudan by using sequencing and molecular tools. We confirmed the presence of An. stephensi in eight border-states, identifying both natural and human-made breeding sites. Our analysis revealed the presence of 20 haplotypes with different distributions per state. This study revealed a countrywide spread of An. stephensi in Sudan, with confirmed presence in borders states with Chad, Egypt, Eritrea, Ethiopia, Libya, Republic of Central Africa, and South Sudan. Detection of An. stephensi at points of entry with these countries, particularly Chad, Libya, and South Sudan, indicates the rapid previously undetected spread of this invasive vector. Our phylogenetic and haplotype analysis suggested local establishment and evolutionary adaptation of the vector to different ecological and environmental conditions in Sudan. Urgent engagement of the global community is essential to control and prevent further spread into Africa.

The role of integrated programs in the prevention of COVID-19 in a humanitarian setting.

The COVID-19 pandemic has caused a severe depletion to health systems worldwide. In the South Kordofan state, a war-torn humanitarian region in Sudan, the United Nations High Commissioner for Refugees and the WHO Emergencies Programme have led the development of integrated programs for health, peace and rehabilitation by training rapid response teams to strengthen both surveillance system and response capacities, engaging the communities at risk with health education and promotion activities, as well as following proper infection, prevention and control measures during case investigation. Also, Early Help Assessment coordinators met with health services managers and recruited 14 teams who were trained to ensure a rapid response to COVID-19. Also, the implementation of water, sanitation and hygiene services was upscaled. Although the local community of South Kordofan is fragile and at high risk of infectious diseases, the limited numbers of detected cases of COVID-19 and COVID-19 deaths could be attributed to the early preparation and integration of programs that helped to prevent the local spread of COVID-19. This lesson needs to be thoroughly investigated to estimate whether it is cost-effective and to determine the feasibility of it being successfully implemented in other humanitarian settings.

Obstacles faced by healthcare providers during COVID-19 pandemic in Sudan.

The impacts of COVID -19 pandemic have been quite significant on healthcare providers. I was particularly challenging for those in Low and Middle-Income Countries including Sudan . Unfortunately, the pandemic has hit Sudan on extremely difficult time for the country and its people. The country was coming out of long-brutal and devastating dictatorship and transitioning to new democracy with civilian leadership. In addition to the pandemic related issues, trying to rebuild the health system during socioeconomic crisis, healthcare providers  in the country were challenged personally and professionally. These challenges include the stress of working in under-resourced settings with limited access to personal-protection equipment and testing kits raised the fear of contracting the virus and spreading it to their families. The professional, social, and personal life of healthcare providers have been dramatically changed by the ongoing pandemic, however, they are heroically accepting this change in a hope that, this will save the life of many more people. Nevertheless, their fights and sacrifices should at least be rewarded by governments and communities altogether strictly enforce the implementation of other preventive measures including vaccination, face masking, and social distancing and get all protected. We should all understand that, unless we are all protected no one is protected, so all must adapt to the new norm of life and collaborate not only on ending this pandemic but to prevent similar ones in the future.

Assessment of genetic diversity of Plasmodium falciparum circumsporozoite protein in Sudan: the RTS,S leading malaria vaccine candidate.

BACKGROUND: The currently used malaria vaccine, RTS,S, is designed based on the Plasmodium falciparum circumsporozoite protein (PfCSP). The pfcsp gene, besides having different polymorphic patterns, can vary between P. falciparum isolates due to geographical origin and host immune response. Such aspects are essential when considering the deployment of the RTS,S vaccine in a certain region. Therefore, this study assessed the genetic diversity of P. falciparum in Sudan based on the pfcsp gene by investigating the diversity at the N-terminal, central repeat, and the C-terminal regions. METHODS: A cross-sectional molecular study was conducted; P. falciparum isolates were collected from different health centres in Khartoum State between January and December 2019. During the study period, a total of 261 febrile patients were recruited. Malaria diagnosis was made by expert microscopists using Giemsa-stained thick and thin blood films. DNA samples were examined by the semi-nested polymerase chain reaction (PCR). Single clonal infection of the confirmed P. falciparum cases, were used to amplify the pfcsp gene. The amplified amplicons of pfcsp have been sequenced using the Sanger dideoxy method. The obtained sequences of pfcsp nucleotide diversity parameters including the numbers of haplotypes (Hap), haplotypes diversity (Hapd), the average number of nucleotide differences between two sequences (p), and the numbers of segregating sites (S) were obtained. The haplotype networks were constructed using the online tcsBU software. Natural selection theory was also tested on pfcsp using Fuand Li's D, Fuand Li's F statistics, and Tajima's D test using DnaSP. RESULTS: In comparison with the different pfcsp reference strains, the Sudanese isolates showed high similarity with other African isolates. The results of the N-terminal region showed the presence of 2 different haplotypes with a Hapd of 0.425 ± 0.00727. The presence of the unique insertion of NNNGDNGREGKDEDKRDGNN was reported. The KLKQP motif was conserved in all the studied isolates. At the central repeat region, 11 haplotypes were seen with a Hapd of 0.779 ± 0.00097. The analysis of the genetic diversity in the C-terminal region showed the presence of 10 haplotypes with a Hapd of 0.457 ± 0.073. Several non-synonymous amino acids changes were also seen at the Th2R and the Th3R T-cell epitope regions including T317K, E317K, Q318E, K321N, I322K, T322K, R322K, K324Q, I327L, G352N, S354P, R355K, N356D, Q357E, and E361A. CONCLUSIONS: In this study, the results indicated a high conservation at the pfcsp gene. This may further contribute in understanding the genetic polymorphisms of P. falciparum prior to the deployment of the RTS,S vaccine in Sudan.