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Abstract Introduction When COVID-19 hit the world in 2019, an enhanced focus on diagnostic testing for SARS-CoV-2 was essential for a successful pandemic response. Testing laboratories stretched their capabilities for the new coronavirus by adopting different test methods. The necessity of having external quality assurance (EQA) mechanisms was even more critical due to this rapid expansion. However, there was a lack of experience in providing the necessary SARS-CoV-2 EQA materials, especially in locations with constrained resources. Objective We aimed to create a PT (Proficiency testing) programme based on the Dried Tube Specimens (DTS) method that would be a practical option for molecular based SARS-CoV-2 EQA in Low- and Middle-Income Countries. Methods Based on previous ISO/IEC 17043:2010 accreditation experiences and with assistance from the US Centers for Disease Control and Prevention, The Supranational Reference Laboratory of Uganda (adapted the DTS sample preparation method and completed a pilot EQA program between 2020 and 2021. Stability and panel validation testing was conducted on the designed materials before shipping to pilot participants in six African countries. Participants received a panel containing five SARS-CoV-2 DTS samples, transported at ambient conditions. Results submitted by participants were compared to validation results. Participants were graded as satisfactory (≥ 80%) or unsatisfactory (

Abstract Background Smear microscopy has remained the initial diagnostic test for presumptive tuberculosis (TB) patients in health facilities without the World Health Organization (WHO) recommended rapid diagnostic tools. In the Uganda TB laboratory network, the technique remains the only tool to monitor response to treatment among drug susceptible TB patients, with the country currently having over 1,600 microscopy TB testing units. It has been evidenced that acid-fast bacilli (AFB) microscopy’s yield highly depends on the staining technique and reading ability of the laboratory personnel. For the quality of TB testing in the country, the TB control program set up a Randomized Blinded Rechecking (RBRC) program in 2008 to monitor the testing performance of laboratories to continuously improve the reliability and efficiency of results. This is the first study to determine the effectiveness and impact of the RBRC program on the performance of the participating laboratories in Uganda. Methods This was a retrospective cross-sectional study based on a record review of the RBRC’s annual results compilations between January 2008 and December 2017. Results Between January 2008 and December 2017, a total of 265,523 smears were re-checked during the RBRC program. The number of enrolled laboratories in the RBRC program rose from 660 to 2008 to 1,406 in 2017. The RBRC program resulted in a statistically significant reduction in microscopy errors, with false positives decreasing from 12.8% to 2008 to 7.6% in 2017, false positive errors decreasing from 10 to 6.3%, false negative errors decreasing from 2.9 to 0.7%, quantification errors decreasing from 6.0 to 1.8%, and the overall sensitivity of smear microscopy compared to the controllers increased with statistical significance from 93 to 97%. Conclusion The study reveals an overall significant error reduction and an improved sensitivity of smear microscopy upon continuous implementation of the RBRC program in an AFB microscopy TB laboratory network. Implementation of a RBRC program is crucial and essential to maintaining a reliable TB laboratory service that can facilitate accurate diagnosis and offset the disadvantages of using smear microscopy.

Abstract Background Mycobacterium tuberculosis presents several lineages each with distinct characteristics of evolutionary status, transmissibility, drug resistance, host interaction, latency, and vaccine efficacy. Whole genome sequencing (WGS) has emerged as a new diagnostic tool to reliably inform the occurrence of phylogenetic lineages of Mycobacterium tuberculosis and examine their relationship with patient demographic characteristics and multidrug-resistance development. Methods 191 Mycobacterium tuberculosis isolates obtained from a 2017/2018 Tanzanian drug resistance survey were sequenced on the Illumina Miseq platform at Supranational Tuberculosis Reference Laboratory in Uganda. Obtained fast-q files were imported into tools for resistance profiling and lineage inference (Kvarq v0.12.2, Mykrobe v0.8.1 and TBprofiler v3.0.5). Additionally for phylogenetic tree construction, RaxML-NG v1.0.3(25) was used to generate a maximum likelihood phylogeny with 800 bootstrap replicates. The resulting trees were plotted, annotated and visualized using ggtree v2.0.4 Results Most [172(90.0%)] of the isolates were from newly treated Pulmonary TB patients. Coinfection with HIV was observed in 33(17.3%) TB patients. Of the 191 isolates, 22(11.5%) were resistant to one or more commonly used first line anti-TB drugs (FLD), 9(4.7%) isolates were MDR-TB while 3(1.6%) were resistant to all the drugs. Of the 24 isolates with any resistance conferring mutations, 13(54.2%) and 10(41.6%) had mutations in genes associated with resistance to INH and RIF respectively. The findings also show four major lineages i.e. Lineage 3[81 (42.4%)], followed by Lineage 4 [74 (38.7%)], the Lineage 1 [23 (12.0%)] and Lineages 2 [13 (6.8%)] circulaing in Tanzania. Conclusion The findings in this study show that Lineage 3 is the most prevalent lineage in Tanzania whereas drug resistant mutations were more frequent among isolates that belonged to Lineage 4.

Abstract Background Uganda remains one of the countries with the highest burden of TB/HIV. Drug-resistant TB remains a substantial challenge to TB control globally and requires new strategic effective control approaches. Drug resistance usually develops due to inadequate management of TB patients including improper treatment regimens and failure to complete the treatment course which may be due to an unstable supply or a lack of access to treatment, as well as patient noncompliance. Methods Two sputa samples were collected from Xpert MTB/RIF® assay-diagnosed multi-drug resistant tuberculosis (MDR-TB) patient at Lira regional referral hospital in northern Uganda between 2020 and 2021 for comprehensive routine mycobacterial species identification and drug susceptibility testing using culture-based methods. Detection of drug resistance-conferring genes was subsequently performed using whole-genome sequencing with Illumina MiSeq platform at the TB Supranational Reference Laboratory in Uganda. Results In both isolates, extensively drug-resistant TB (XDR-TB) was identified including resistance to Isoniazid (katG p.Ser315Thr), Rifampicin (rpoB p.Ser450Leu), Moxifloxacin (gyrA p.Asp94Gly), Bedaquiline (Rv0678 Glu49fs), Clofazimine (Rv0678 Glu49fs), Linezolid (rplC Cys154Arg), and Ethionamide (ethA c.477del). Further analysis of these two high quality genomes revealed that this 32 years-old patient was infected with the Latin American Mediterranean TB strain (LAM). Conclusions This is the first identification of extensively drug-resistant Mycobacterium tuberculosis clinical isolates with bedaquiline, linezolid and clofazimine resistance from Uganda. These acquired resistances were because of non-adherence as seen in the patient’s clinical history. Our study also strongly highlights the importance of combating DR-TB in Africa through implementing next generation sequencing that can test resistance to all drugs while providing a faster turnaround time. This can facilitate timely clinical decisions in managing MDR-TB patients with non-adherence or lost to follow-up.

BackgroundProficiency testing (PT) has been hard to set up due to cost limitations and technical capacity. Conventional Xpert MTB/RIF PT programs use liquid and culture spots which require stringent storage and transportation conditions with cross-contamination chances prevalent. These setbacks prompted the use of dried tube specimens (DTS) for Ultra assay PT. For continuity of PT provision, stability of DTS and compatibility with testing protocols when kept for a long period needs to be established.MethodsDTS were prepared from known isolates inactivated using a hot air oven at 85°C. 100μl of bacterial suspensions were aliquoted and dried inside a Biosafety cabinet. Panel validation was done to establish the baseline Deoxyribonucleic acid (DNA) concentration in terms of cycle threshold (Ct) value. DTS aliquots were shipped to participants to test and report within six weeks. The remaining DTS were kept at 2-8°C and room temperature for one year with testing at six months. Twenty (20) DTS samples per set remaining at one year were heated at 55°C for two weeks before testing. The means of the different samples were compared to validation data using paired t-tests. Boxplots were designed to visualize the differences in the medians of the DTS.ResultsOverall mean Ct value increased by 4.4 from the validation to testing after one year at the different storage conditions. Samples heated at 55°C showed a 6.4 Ct difference from validation data. Testing done at six months on 2-8°C stored items showed no statistical difference. At all the remaining testing times and conditions, P-values were less than 0.008 although the absolute mean Ct when compared showed slight increments and accommodated differences for the detection of MTB and rifampicin resistance. Median values for samples stored at 2-8°C were lower compared to those at room temperature.ConclusionDTS stored at 2-8°C remain more stable for one year compared to higher temperatures and can be consistently used as PT materials in more than one PT round for biannual PT providers.

Corporate e-learning refers to the use of online learning technologies to provide professional training to employees. E-learning offers many benefits for organizations, including cost savings, flexibility, and provide consistent, up-to-date training to a diverse workforce. Implementing corporate e-learning programs can improve corporate learning, but also presents challenges such as developing effective content, engaging learners, and measuring the impact of the training. This study was conducted as a systematic literature review to find the missing gaps in corporate e-learning research by collecting past research articles from 4 scientific research databases in the last decade. In total 7,285 records were collected across the 4 databases which went through the PRISMA and rigorous process of inclusion and exclusion, and selection processes. In conclusion, 21 research articles were chosen for inclusion in the final review. This SLR found that e-learning has become increasingly popular in corporate learning over the past 15 years despite challenges in implementation, also offering benefits such as cost reduction, ease of access, comfort, learning ability, and speed, which improves productivity and efficiency in organization. Hopefully, the results of this SLR will benefit researchers who seek insight in the field of corporate e-learning and assist organizations to make informed decisions in deploying or improving e-learning in their establishments. [ABSTRACT FROM AUTHOR]

Abstract Background Diabetic peripheral neuropathy (DPN), due to its potential for causing morbidity and disability from foot ulcers and amputations, is increasingly becoming a source of concern in Saudi Arabia and worldwide. However, wide variability exists in the prevalence of DPN reported in previous studies in Saudi Arabia, limiting the utility of existing data in national public health policy. Therefore, the aim of this study was to systematically evaluate the magnitude of DPN in patients living with DM in Saudi Arabia in order to inform policymakers during the implementation of appropriate preventive and treatment strategies for DPN. Methods PubMed, Google Scholar, African Journals Online, Scopus, Web of Science, Embase, and Wiley Online Library were searched systematically to acquire relevant articles based on preset criteria. We evaluated heterogeneity and publication bias and employed a random-effects model to estimate the pooled prevalence of DPN from the included studies. We followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines in conducting the meta-analysis. Analysis was performed using the STATA Version 12 software. Results Twelve studies with a total of 4,556 participants living with DM, of whom 2,081 were identified as having DPN were included in the meta-analysis. The overall prevalence of DPN was 39% (95% CI [30%, 49%]). Subgroup analysis based on diagnostic method showed that prevalence estimates for DPN using screening questionnaires and clinical examination were 48% (95% CI [46%, 50%]) and 40% (95% CI: [38%, 42%]), respectively, while the estimated prevalence using nerve conduction studies was 26% (95% CI [15%, 36%]). Conclusion This study showed a high magnitude of DPN in Saudi Arabia (39%), thus highlighting the need for sustained efforts to reduce the prevalence of diabetes mellitus and DPN in the kingdom.

Objective: The study was carried out to analyze the various phytochemical properties and to determine the in-vitro antimicrobial activity of Parkia biglobosa methanolic leaf extract against Methicillin-Resistant Staphylococcus aureus (MRSA). Materials and methods: The phytochemical screening and Thin Layer Chromatography (TLC) of the methanol extract were conducted using standard methods. The antimicrobial activity of the P. biglobosa methanolic leaf extract on sheep and human MRSA isolates was investigated by disc diffusion method. The extract was tested at different concentrations of 100, 200 and 400 mg/mL and norfloxacin (10 µg) as control. The MIC/MBC were determined by serial dilution method in broth media. Results: the phytochemical analysis showed the presence of alkaloid, saponins, tannins, cardiac glycosides, flavonoids and terpenoids while TLC result revealed 5 spots with retention factor (Rf) values of 0.14, 0.34, 0.42, 0.56 and 0.66 depicting different component using methanol and chloroform at the ratio of 4:1 as solvent system. The diameter of inhibition zone (DIZ) of the extract on MRSA isolates cultured on agar plate revealed a dose dependent response. The extract had the highest DIZ average replicate value of 20.66±1.76 against human MRSA isolates and 20.66±1.20 was recorded as highest DIZ on the sheep MRSA isolates. Norfloxacin had the highest DIZ of 20.33±0.57 on sheep MRSA isolates and 20.33±1.52 DIZ against human isolates. MIC/MBC revealed the highest values of 6.25 mg/mL/12.5 mg/mL for both sheep and human MRSA isolates. Conclusion: The susceptibility test of crude methanol extract of P. biglobosa leaf has revealed activity against MRSA. Thereby, attesting to the folkloric claim of using P. biglobosa for ethno-therapeutic purposes against infectious diseases in Nigeria and other places. [J Adv Vet Anim Res 2018; 5(2.000): 173-181]

Aim: Coccidiosis is an important enteric parasitic disease of poultry associated with significant economic losses to poultry farmers worldwide. This survey was conducted from June 2014 through July 2015 with the main goal of investigating the prevalence and associated risk factors of coccidiosis among village and exotic breeds of chickens in Maiduguri, Northeastern Nigeria. Materials and Methods: A total of 600 fecal samples from live and slaughtered birds comprising 284 young, 141, growers and 175 adult birds; 379 male and 221 female birds; 450 exotic and 150 local breeds of birds were randomly collected either as bird’s fresh droppings or cutting open an eviscerated intestine of slaughtered birds, while noting their age, sex, and breeds. Samples were analyzed using standard parasitological methods and techniques. Results: An overall prevalence rate of 31.8% (95% confidence interval: 28.07-35.52) was obtained. Higher prevalence rates were recorded in growing birds 58.9% (50.78-67.02), female birds 35.3% (29.00-41.60), exotic birds 42.4% (37.83- 46.97), and broiler birds 68.7% (61.28-76.12). Similarly, higher infection rates were also observed among birds sampled from Mairi ward 66.7% (56.03-77.37), intensive management system 46.5% (41.61-51.39), and constructed local cages 54.0% (46.02-61.98). The difference in prevalence of coccidiosis among age groups, breeds, among exotic breeds, sampling locations, husbandry management systems, and litter management systems was statistically significant (0.05) of infection rates was observed in sex. Conclusion: Coccidiosis is endemic in both commercial and backyard poultry farms in Maiduguri due to poor management practices encouraging Eimeria oocysts build-up. It is therefore, recommended that poultry farmers should practice strict biosecurity measures on their farms, creating awareness on the prevalence of coccidiosis, routine vaccination against coccidiosis and educating poultry farmers on the need for maintaining good hygienic standards and good flock health management.

Objective: The objectives of this study was to investigate phenotypically the prevalence of Staphylococcus aureus and Methicillin Resistance S. aureus (MRSA) in ruminants and animal handlers in Maiduguri, Nigeria. Materials and methods: A total of 937 samples (900 nasal and skin swabs of cattle and 37 humans samples) were collected in Maiduguri metropolis. The samples were inoculated onto mannitol salt agar (MSA) and blood agar, and the cultrue plates were incubated at 37oC for 24 h. The S. aureus colonies showing golden yellow color on MSA were primarily identified as S. aureus, which were then subjected for catalase and coagulase tests. All S. aureus were finally screened for the presence of MRSA on oxalate resistant screening agar (ORSAB) medium. Antimicrobial susceptibility of the MRSA were measured by disc diffusion method. Results: The overall prevalence of S. aureus was 44.3% (n=414/937). The MRSA could be detected in 12.1% (n=113/937) samples. Considerign different species, S. aureus could be isolated from 137 (45.6%), 148 (49.3%) and 119 (39.7%), 11 (29.7%) cattle, sheep, goat and humans, respectively. In ruminants, 27.7% (n-83/300) bulls were positive for S. aureus. Similarly, 10.7% (n=32/300) and 22.3% (n=67/300) were positive for S. aureus in rams and bucks, respectively. On the other hand, 18% (n=54/300), 38.7% (n=116/300) and 17.3% (n=52/300) samples from cows, ewes and does were positive for S. aureus. The highest MRSA could be isolated from ewes (9.3%; n=28/300) follwed by bulls (7%; n=21/300) and bucks (6.7%; n=20/300). In humans, 13.5% (n=5/37) samples were positive for MRSA. Conclusion: S. aureus and MRSA infections in ruminants and animal handlers are documented in this study. The MRSA may exert public health threat to humans. [J Adv Vet Anim Res 2016; 3(2.000): 152-159]

Tick-borne diseases (TBDs) or otherwise called equine piroplasmosis (EP) are the foremost economic limitations to equids production. Thus, reducing the breeding capability and athletic performance of equids globally. Identification of these haemoparasites is crucial in understanding their distribution in the population and it is imperative to discern between species and subspecies that are responsible for the occurrence of the disease conditions. Conventional procedures such as microscopic and serological evaluations do not usually meet these prerequisites. Diagnostic contrivances, such as the complement fixation test (CFT), the indirect fluorescent antibody test (IFAT) and the enzyme linked immunosorbent assay (ELISA) have been efficaciously used for many years. Furthermore, DNA-based investigations for identification, differentiation and classification of different haemoparasites have also been established. Molecular diagnostic procedures, such as DNA hybridization, polymerase chain reaction (PCR), transcriptomics, proteomics, metagenomics and metabolomics, permit the uncovering of parasites in blood, tissues or ticks with optimal sensitivity, specificity and consistency. In addition, these procedures can be exploited to detect definite species and subspecies. The prerequisite of these investigations must include proper premeditation and validation, these investigations provide an effective device for molecular studies, with greater benefits of flexibility to standardization. The application of these procedures for studying TBDs or EP globally will be irreplaceable for a long period from now. Therefore, the aim of this review is to draw up the specifics of the procedures in more convenient form for practitioners and researchers. KEY WORDS: Diagnosis, equids, molecular, transcriptomics, proteomics, metagenomics, metabolomics, haemoparasites, tick-borne diseases [J Adv Vet Anim Res 2016; 3(2.000): 84-91]

Aim: To determine the presence of MRSA in hunting dogs in Maiduguri metropolitan. Materials and Methods: Phenotypic methods used includes microscopic technique, colony morphology study, catalase-coagulase tests, and the use of mannitol salt agar test, oxacillin resistance screening agar base, and antibiotic susceptibility testing methods. Genotypic approach was used for deoxyribonucleic acid extraction, and the presence of nuc and mecA gene was detected using polymerase chain reaction (PCR) techniques. Results: Examination of 416 swab samples from nasal and perineal region of dogs revealed a total of 79.5% of S. aureus, where 62.5% of the isolates were MRSA. Molecular analysis revealed that 7nuc genes specific for S. aureus from 20 presumptive MRSA assay were all mecA PCR negative. The isolates were sensitive to gentamicin and ciprofloxacin but proved resistant to cefoxitin and oxacillin. Conclusion: High isolation rate of MRSA was found in hunting dogs. Significant level (p