Cellular and Cytokine Responses in the Granulomas of Asymptomatic Cattle naturally infected with Mycobacterium bovis in Ethiopia

Cells (CD3+ T cell and CD68+ macrophages), cytokines (IFN-γ+ and TNF-α+) and effector molecule (iNOS+) responses were evaluated in the lymph nodes and tissue of cattle naturally infected with Detailed post mortem and immunohistochemical examinations of lesions were performed on 16 cows positive for single intradermal cervical comparative tuberculin (SICCT) test which were identified from dairy farms located around the Addis Ababa City. The severity of the gross lesion was significantly higher (p=0.003) in culture positive (n=12) cows than in culture negative (n=4). Immunohistochemical techniques showed that in culture positive cows, the mean immunolabeling fraction of CD3+ T cells decreased as the stage of granuloma increased from stage I to stage IV (p<0.001). In contrast, the immunolabelling fraction of CD68+ macrophages, IFN-γ+, TNF-α+ and iNOS+ increased from stage I to stage IV (p< 0.001). In culture negative cows, early stages showed a significantly higher fraction of CD68+ macrophages (p=0.03) and iNOS+ (p=0.007) when compared to culture positive cows. Similarly, at advanced granuloma stages, culture negative cows demonstrated significantly higher mean proportions of CD3+ T cells (p< 0.001) compared to culture positive cows. Thus, this study demonstrates that following natural infection of cows with , as the stage of granuloma increases from stage I to stage IV, the immunolabelling fraction of CD3+ cells decreases while the immunolabeling fraction of CD68+ macrophages, IFN-γ+, TNF-α+ and iNOS+ increases. [Abstract copyright: Copyright © 2020 Tulu et al.

Epidemiology of Mycobacterium tuberculosis lineages and strain clustering within urban and peri-urban settings in Ethiopia

Background Previous work has shown differential predominance of certain Mycobacterium tuberculosis (M. tb) lineages and sub-lineages among different human populations in diverse geographic regions of Ethiopia. Nevertheless, how strain diversity is evolving under the ongoing rapid socio-economic and environmental changes is poorly understood. The present study investigated factors associated with M. tb lineage predominance and rate of strain clustering within urban and peri-urban settings in Ethiopia. Methods Pulmonary Tuberculosis (PTB) and Cervical tuberculous lymphadenitis (TBLN) patients who visited selected health facilities were recruited in the years of 2016 and 2017. A total of 258 M. tb isolates identified from 163 sputa and 95 fine-needle aspirates (FNA) were characterized by spoligotyping and compared with international M.tb spoligotyping patterns registered at the SITVIT2 databases. The molecular data were linked with clinical and demographic data of the patients for further statistical analysis. Results From a total of 258 M. tb isolates, 84 distinct spoligotype patterns that included 58 known Shared International Type (SIT) patterns and 26 new or orphan patterns were identified. The majority of strains belonged to two major M. tb lineages, L3 (35.7%) and L4 (61.6%). The observed high percentage of isolates with shared patterns (n = 200/258) suggested a substantial rate of overall clustering (77.5%). After adjusting for the effect of geographical variations, clustering rate was significantly lower among individuals co-infected with HIV and other concomitant chronic disease. Compared to L4, the adjusted odds ratio and 95% confidence interval (AOR; 95% CI) indicated that infections with L3 M. tb strains were more likely to be associated with TBLN [3.47 (1.45, 8.29)] and TB-HIV co-infection [2.84 (1.61, 5.55)]. Conclusion Despite the observed difference in strain diversity and geographical distribution of M. tb lineages, compared to earlier studies in Ethiopia, the overall rate of strain clustering suggests higher transmission and warrant more detailed investigations into the molecular epidemiology of TB and related factors

Prevalence of bovine tuberculosis and its associated risk factors in the emerging dairy belts of regional cities in Ethiopia

Bovine tuberculosis (BTB) has become an economically important disease in dairy herds found in and around Addis Ababa City and is emerging in regional cities like Gondar, Hawassa and Mekelle because of the establishment of dairy farms in the milk sheds of these cities. A cross-sectional study to estimate the prevalence of BTB and identify associated risk factors was conducted between February 2016 and March 2017. A total of 174 herds comprising of 2,754 dairy cattle in the cities of Gondar, Hawassa and Mekelle were tested using the Single Intradermal Comparative Cervical Tuberculin (SICCT) test. Data on herd structure, animal origin, body condition, housing condition, farm hygiene, management and biosecurity practices were collected using a pre-tested structured questionnaire. Generalized Linear Models (GLM) and Generalized Linear Mixed Models (GLMM) were used to analyze the herd and animal level risk factors, respectively. The herd prevalence was 22.4% (95% CI: 17–29%) while the animal prevalence was 5.2% (95% CI: 4–6%) at the cut-off >4 mm. The herd prevalence rose to 65.5% (95% CI: 58–72%) and the animal prevalence rose to 9% (95% CI: 8–10%) when the severe interpretation of >2 mm cut-off was applied. The mean within-herd prevalence in positive farms at the cut-off >4 mm was 22.7% (95% CI: 15–31%). At the herd level, the analysis showed that herd size, farm hygiene, feeding condition and biosecurity were significantly associated with BTB status, while new cattle introductions showed only borderline significance and that age of farm, housing condition, farmers’ educational status and animal health care practice were not significant. At the animal level, the results showed that age and animal origin were identified as significant predictors for BTB positivity but sex and body condition score were not related to BTB status. Descriptive analysis revealed that herds having ‘BTB history’ showed slightly higher likelihood of being BTB positive compared to farms having no previous BTB exposure. In conclusion, this study showed relatively lower average prevalence in the emerging dairy regions as compared to the prevalence observed in and around Addis Ababa City, warranting for implementation of control program at this stage to reduce or possibly stop further transmission of BTB

Network analysis of dairy cattle movement and associations with bovine tuberculosis spread and control in emerging dairy belts of Ethiopia

Background: Dairy cattle movement could be a major risk factor for the spread of bovine tuberculosis (BTB) in emerging dairy belts of Ethiopia. Dairy cattle may be moved between farms over long distances, and hence understanding the route and frequency of the movements is essential to establish the pattern of spread of BTB between farms, which could ultimately help to inform policy makers to design cost effective control strategies. The objective of this study was, therefore, to investigate the network structure of dairy cattle movement and its influence on the transmission and prevalence of BTB in three emerging areas among the Ethiopian dairy belts, namely the cities of Hawassa, Gondar and Mekelle. Methods: A questionnaire survey was conducted in 278 farms to collect data on the pattern of dairy cattle movement for the last 5 years (September 2013 to August 2018). Visualization of the network structure and analysis of the relationship between the network patterns and the prevalence of BTB in these regions were made using social network analysis. Results: The cattle movement network structure display both scale free and small world properties implying local clustering with fewer farms being highly connected, at higher risk of infection, with the potential to act as super spreaders of BTB if infected. Farms having a history of cattle movements onto the herds were more likely to be affected by BTB (OR: 2.2) compared to farms not having a link history. Euclidean distance between farms and the batch size of animals moved on were positively correlated with prevalence of BTB. On the other hand, farms having one or more outgoing cattle showed a decrease on the likelihood of BTB infection (OR = 0.57) compared to farms which maintained their cattle. Conclusion: This study showed that the patterns of cattle movement and size of animal moved between farms contributed to the potential for BTB transmission. The few farms with the bulk of transmission potential could be efficiently targeted by control measures aimed at reducing the spread of BTB. The network structure described can also provide the starting point to build and estimate dynamic transmission models for BTB, and other infectious disease

Spoligotyping and drug sensitivity of Mycobacterium tuberculosis isolated from pulmonary tuberculosis patients in the Arsi Zone of southeastern Ethiopia

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Beliefs About Body Weight and Practices of Regulating Food and Physical Exercise Patterns of Children Among Parents in Southern Ethiopia

Bewunetu Zewude,1 Getahun Siraw,2 Belayneh Melese,3 Tewodros Habtegiorgis,1 Ashenafi Hizkeal,1 Mihret Tadele1 1Department of Sociology, College of Social Sciences and Humanities, Wolaita Sodo University, Wolaita Sodo town, Ethiopia; 2Department of Sociology, College of Social Sciences and Humanities, Dilla University, Dilla town, Ethiopia; 3Department of Civics and Ethical Studies, College of Social Sciences and Humanities, Wolaita Sodo University, Wolaita Sodo town, EthiopiaCorrespondence: Bewunetu Zewude, Email [email protected]: The problem of childhood obesity is often associated with cultural values of body weight which includes the belief that voluptuous body size is a sign of good health status and better parental care. We examined the beliefs and behaviors of parents pertaining to preferred body weight of children, including practices of management of feeding and physical exercise patterns of children in southern Ethiopia.Methods: Using cross-sectional survey study design, a structured questionnaire was distributed to randomly selected parents. Quantitative data were collected from 215 respondents, inserted into SPSS software, and analyzed by frequency and percentage distributions, charts, and multinomial logistic regression techniques.Results: Most (59.1%) respondents want their child’s body weight to be normal/neither fat nor thin. In addition, 58.2% of the study participants held the opinion that average body size is an indication of good health status. Furthermore, 72.6% of respondents revealed that they control the amount of food their child consumes with the aim of preventing obesity/overweight. Moreover, 79.5% of respondents encourage their children to sometimes walk on foot, instead of always relying on vehicles while 87% of them replied that they regulate the amount of time their children spend watching TV. Above all, parents’ preference of voluptuous body size of children is significantly associated with age (OR=61.52.448, P < 0.001, 95%C.I: 57– 66.14), educational status (OR = 35.605, P < 0.001; 95%C.I: 6.050– 209.524), sex (OR = 0.237, P < 0.05; 95%C.I: 0.078-0.724) and grownup area (OR = 2.869, P < 0.05; 95%C.I: 1.062– 7.749).Conclusion: Parents not only have adequate level of awareness regarding the health risks of children’s heavy body weight but also engage in the body weight management practices of their children such as by regulating the food consumption patterns of children, encouraging them to undertake physical exercises, and by being role models while doing physical exercises for themselves.Keywords: childhood obesity, body weight, food consumption, physical exercise, regulatio

Equine Histoplasmosis in Ethiopia: Phylogenetic analysis by sequencing of the internal transcribed spacer region of rRNA genes

Equine histoplasmosis commonly known as epizootic lymphangitis (EL) is a neglected granulomatous disease of equine that is endemic to Ethiopia. It is caused by Histoplasma capsulatum variety farciminosum, a dimorphic fungus that is closely related to H. capsulatum variety capsulatum. The objective of this study was to undertake a phylogenetic analysis of H. capsulatum isolated from EL cases of horses in central Ethiopia and evaluate their relationship with H. capsulatum isolates in other countries and/or clades using the internal transcribed spacer (ITS) region of rRNA genes. Clinical and mycological examinations, DNA extraction, polymerase chain reaction (PCR), Sanger sequencing, and phylogenetic analysis were used for undertaking this study. Additionally, sequence data of Histoplasma isolates were retrieved from GenBank and included for a comprehensive phylogenetic analysis. A total of 390 horses were screened for EL and 97 were positive clinically while H. capsulatum was isolated from 60 horses and further confirmed with PCR, of which 54 were sequenced. BLAST analysis of these 54 isolates identified 29 H. capsulatum isolates and 14 isolates from other fungal genera while the remaining 11 samples were deemed insufficient for further downstream analysis. The phylogenetic analysis identified five clades, namely, African, Eurasian, North American 1 and 2, and Latin American A and B. The Ethiopian isolates were closely aggregated with isolates of the Latin American A and Eurasian clades, whereas being distantly related to isolates from North American 1 and 2 clades as well as Latin American B clade. This study highlights the possible origins and transmission routes of Histoplasmosis in Ethiopia

Detection of <i>M. tuberculosis</i> DNA in CD34-Positive Peripheral Blood Mononuclear Cells of Asymptomatic TB Contacts

Haematopoietic stem cells expressing the CD34 surface marker have been posited as a niche for Mycobacterium tuberculosis complex bacilli during latent tuberculosis infection. Our aim was to determine whether M tuberculosis complex DNA is detectable in CD34-positive peripheral blood mononuclear cells (PBMCs) isolated from asymptomatic adults living in a setting with a high tuberculosis burden

Detection of Mycobacterium tuberculosis complex DNA in CD34-positive peripheral blood mononuclear cells of asymptomatic tuberculosis contacts: an observational study

Background: Haematopoietic stem cells expressing the CD34 surface marker have been posited as a niche for Mycobacterium tuberculosis complex bacilli during latent tuberculosis infection. Our aim was to determine whether M tuberculosis complex DNA is detectable in CD34-positive peripheral blood mononuclear cells (PBMCs) isolated from asymptomatic adults living in a setting with a high tuberculosis burden. Methods: We did a cross-sectional study in Ethiopia between Nov 22, 2017, and Jan 10, 2019. Digital PCR (dPCR) was used to determine whether M tuberculosis complex DNA was detectable in PBMCs isolated from 100 mL blood taken from asymptomatic adults with HIV infection or a history of recent household or occupational exposure to an index case of human or bovine tuberculosis. Participants were recruited from HIV clinics, tuberculosis clinics, and cattle farms in and around Addis Ababa. A nested prospective study was done in a subset of HIV-infected individuals to evaluate whether administration of isoniazid preventive therapy was effective in clearing M tuberculosis complex DNA from PBMCs. Follow-up was done between July 20, 2018, and Feb 13, 2019. QuantiFERON-TB Gold assays were also done on all baseline and follow-up samples. Findings: Valid dPCR data (ie, droplet counts >10 000 per well) were available for paired CD34-positive and CD34-negative PBMC fractions from 197 (70%) of 284 participants who contributed data to cross-sectional analyses. M tuberculosis complex DNA was detected in PBMCs of 156 of 197 participants with valid dPCR data (79%, 95% CI 74–85). It was more commonly present in CD34-positive than in CD34-negative fractions (154 [73%] of 197 vs 46 [23%] of 197; p<0·0001). Prevalence of dPCR-detected M tuberculosis complex DNA did not differ between QuantiFERON-negative and QuantiFERON-positive participants (77 [78%] of 99 vs 79 [81%] of 98; p=0·73), but it was higher in HIV-infected than in HIV-uninfected participants (67 [89%] of 75 vs 89 [73%] of 122, p=0·0065). By contrast, the proportion of QuantiFERON-positive participants was lower in HIV-infected than in HIV-uninfected participants (25 [33%] of 75 vs 73 [60%] of 122; p<0·0001). Administration of isoniazid preventive therapy reduced the prevalence of dPCR-detected M tuberculosis complex DNA from 41 (95%) of 43 HIV-infected individuals at baseline to 23 (53%) of 43 after treatment (p<0·0001), but it did not affect the prevalence of QuantiFERON positivity (17 [40%] of 43 at baseline vs 13 [30%] of 43 after treatment; p=0·13). Interpretation: We report a novel molecular microbiological biomarker of latent tuberculosis infection with properties that are distinct from those of a commercial interferon-γ release assay. Our findings implicate the bone marrow as a niche for M tuberculosis in latently infected individuals. Detection of M tuberculosis complex DNA in PBMCs has potential applications in the diagnosis of latent tuberculosis infection, in monitoring response to preventive therapy, and as an outcome measure in clinical trials of interventions to prevent or treat latent tuberculosis infection. Funding: UK Medical Research Council