Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

BACKGROUND: In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains, posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. METHODS: A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. RESULTS: S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. CONCLUSION: This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

Factors affecting the microbiological quality and contamination of farm bulk milk by Staphylococcus aureus in dairy farms in Asella, Ethiopia.

BACKGROUND: The determination of the microbiological quality and safety of raw milk and the associated influencing factors at the farm level is very critical given that the quality or safety of subsequent products that are further produced depends on this. Therefore, this study aimed to determine the microbiological quality and safety of bulk milk and identify associated risk factors, and assess the presence/absence of S. aureus in bulk milk with potential contaminating sources in dairy farms in Asella, Ethiopia. RESULTS: The geometric means of bacterial counts in farm bulk milk were 5.25 log cfu/ml, 3.1 log cfu/ml and 2.97 log cfu/ml for total bacterial count (TBC), coliform count (CC) and coagulase-positive staphylococci count (CPS), respectively. Of the 50 dairy farms, 66, 88, and 32% had TBC, CC and CPS counts, respectively, that exceeded the standard international limits for raw cow's milk intended for direct human consumption. TBC tended to increase as CC increased in bulk milk (r = 0.5). In the final regression model, increased TBC, CC and the contamination of farm bulk milk by S. aureus were significantly associated with dirty barns, dirty cows and soiled udder and teats. TBC was higher during the rainy season than during the dry season. The reported practice of washing teats with warm water significantly decreased CC and CPS. The occurrence of S. aureus was significantly (p < 0.05) higher in bulk farm milk (42%) than in pooled udder milk (37.3%), teat swabs (22.5%), milkers' hand swabs (18%), bulking bucket swabs (16.7%), milking container swabs (14%), and water for cleaning of udder and milkers' hands (10%). The questionnaire survey result showed widespred raw milk consumption habits, low level of training and poor hygienic milking practices. CONCLUSIONS: This study revealed low-quality bulk farm milk with high bacterial counts and a high occurrence of S. aureus. This indicates the potential food safety risks due to consumption of raw milk or its products. This study suggests awareness creation to dairy farmers and the public on hygienic milk production and heat treatment of milk before consumption.

Prevalence of paratuberculosis in cattle based on gross and microscopic lesions in Ethiopia.

BACKGROUND: Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic progressive granulomatous enteritis mainly affecting domestic and wild ruminants worldwide. Although paratuberculosis could be prevail in Ethiopia, there is a scarcity of epidemiological data on paratuberculosis in the country. Thus, this study was conducted to estimate the prevalence of paratuberculosis based on gross and microscopic lesions in cattle slaughtered at ELFORA Abattoir, central Ethiopia. Small intestines and associated lymph nodes of 400 apparently healthy cattle which were slaughtered at ELFORA export abattoir were examined for gross and microscopic lesions of paratuberculosis. The microscopic lesions were classified into four grades (I-IV) based on the type and number of cells infiltrated into the lesion. The prevalence of paratuberculosis was estimated on the basis of gross as well as microscopic lesion of paratuberculosis. RESULTS: The prevalence of paratuberculosis was 11.25% (95% Confidence interval, CI = 0.083-0.148) on the basis of gross lesion. However, relatively lower prevalence (2.0%, 95% CI = 0.01, 0.039) was recorded based on microscopic lesion. The gross lesions were characterized by intestinal thickening, mucosal corrugations and enlargement of associated mesenteric lymph nodes. On the other hand, the microscopic lesions were characterized by granuloma of different grades ranging from grade I to grade III lesions. CONCLUSIONS: The present study indicated the occurrence of paratuberculosis in cattle of Ethiopia based on the detection of gross and microscopic lesions consistent with the lesion of paratuberculosis. The result of this study could be used as baseline information for future studies on the epidemiology and economic significance of paratuberculosis.

Isolation of nematophagous fungi from soil samples collected from three different agro-ecologies of Ethiopia.

BACKGROUND: Several species of nematophagous fungi exist in nature that can capture and kill nematodes as natural predators of soil-dwelling worms. These are important in agriculture and animal husbandry as biological control agents. The diversity of nematophagous fungi found from soil had not been studied in Ethiopia. OBJECTIVE: This study aimed to isolate Nematophagous Fungi from Soil Samples Collected From three Different Agro-Ecologies of Ethiopia. METHODS: Cross-sectional study was conducted and samples were collected from three different agro-climatic zones of Ethiopia; Debre-Berhan (highland), Bishoftu (mid-altitude), and Awash (lowland). Twenty-seven soil samples were randomly taken from each of the three different agro-ecological climates (9 from each agro-ecological climatic zone). For each study site, samples were collected from the soil of decomposed animal feces/dung, agricultural/farmlands, and forest lands in triplicates. RESULTS: The present study disclosed that nematophagous fungi were widespread from the study area. A total of 33 species of nematophagous fungi belonging to four genera, Arthrobotryes, Paecilomyces, Monacrosporium, and Harposporium were identified. Arthrobotrys were the most commonly isolated genera followed by Paecilomyces. The six identified species were Arthrobotrys oligospora, Paecilomyces lilacinus, Arthrobotryes dactyloides, Monacosporum eudermatum, Harposporium helicoides, and Monacosporum cionopagum. CONCLUSION: This study indicated that Arthrobothryes oligospora was the most common species in Bishoftu and Awash whereas. In Debre-Berhan, Paecilomyces lilacinus was the most prevalent species. Monacosporum cionapagum was not isolated from dung soil and agricultural soil whereas Harposporium helicoides and Arthrobothryes dactyloides were not found from dung and forest soil respectively.

Transcriptional responses of Daphnia magna exposed to Akaki river water.

Pollution of the aquatic environment is a global problem, with industrial waste, farming effluents, sewage, and wastewater as the main contributors. Many pollutants are biologically active at low concentrations, resulting in sublethal effects, which makes it a highly complex situation and difficult to assess. In many places, such as the Akaki river in Ethiopia, the pollution situation has resulted in streams with minimal presence of invertebrates or vertebrates. As it is difficult to perform a complete chemical analysis of the waters, the present study focused on using gene expression analysis as a biological end point to determine the effects of Akaki river contaminants. The present study was conducted using the small planktonic crustacean Daphnia magna with toxicogenomic molecular markers. Daphnia magna neonates were exposed to Akaki water samples collected from two different sites on the river and analyzed for mortality and expression of genes involved in different biological pathways. Despite the poor quality of Akaki river water, 48 h acute toxicity tests showed no mortality. Interestingly, analysis of sublethal toxicogenomic responses showed that exposure to Akaki water altered the expression of 25 out of 37 genes involved in metal regulation, immune response, oxidative stress, respiration, reproduction, and development. The toxicogenomic data gives insight into the mechanisms involved in causing potential adverse effects to aquatic biota harboring the Akaki river system.

MERS coronaviruses from camels in Africa exhibit region-dependent genetic diversity.

Middle East respiratory syndrome coronavirus (MERS-CoV) causes a zoonotic respiratory disease of global public health concern, and dromedary camels are the only proven source of zoonotic infection. Although MERS-CoV infection is ubiquitous in dromedaries across Africa as well as in the Arabian Peninsula, zoonotic disease appears confined to the Arabian Peninsula. MERS-CoVs from Africa have hitherto been poorly studied. We genetically and phenotypically characterized MERS-CoV from dromedaries sampled in Morocco, Burkina Faso, Nigeria, and Ethiopia. Viruses from Africa (clade C) are phylogenetically distinct from contemporary viruses from the Arabian Peninsula (clades A and B) but remain antigenically similar in microneutralization tests. Viruses from West (Nigeria, Burkina Faso) and North (Morocco) Africa form a subclade, C1, that shares clade-defining genetic signatures including deletions in the accessory gene ORF4b Compared with human and camel MERS-CoV from Saudi Arabia, virus isolates from Burkina Faso (BF785) and Nigeria (Nig1657) had lower virus replication competence in Calu-3 cells and in ex vivo cultures of human bronchus and lung. BF785 replicated to lower titer in lungs of human DPP4-transduced mice. A reverse genetics-derived recombinant MERS-CoV (EMC) lacking ORF4b elicited higher type I and III IFN responses than the isogenic EMC virus in Calu-3 cells. However, ORF4b deletions may not be the major determinant of the reduced replication competence of BF785 and Nig1657. Genetic and phenotypic differences in West African viruses may be relevant to zoonotic potential. There is an urgent need for studies of MERS-CoV at the animal-human interface.

Towards objective measurement of reproductive performance of traditionally managed goat flocks in the drylands of Ethiopia.

Reproductive performance is a key determinant for the efficiency of goat production. Regular monitoring of reproductive efficiency is essential to assess management and to avoid financial losses due to poor performance. To allow more objective measurement and comparisons over time, we propose a novel quantitative approach for defining annual reproductive performance by combining common performance indicators into a goat flock index. Commonly used reproductive performance measures were collected from 242 goat flocks in four districts in dryland of Ethiopia between July 2018 and February 2019. Principal component analysis (PCA) was performed to identify biologically meaningful latent components that explain annual reproductive output (ARO) and annual reproductive wastage (ARW). Together with the remaining annual reproductive performance measures, the ARO and ARW components were included in a PCA to derive an algorithm for a goat annual reproductive performance index (G-ARPI). One component representing variation in kidding interval, PCARO1 and PCARW1 was extracted and normalized to a 10-scale value. The flocks were classified into good performing (15.63%) with index > 8.5, moderately performing (48.21%) with index values ranging from 6.5 to 8.5 and poor performing (36.16%) with index < 6.5. Good performing flocks have higher scores for reproductive output measures, lower scores for reproductive wastage and lower kidding interval. The proposed G-ARPI can be used as an objective tool to compare reproductive performance between management systems, evaluate the costs of poor reproductive management and will be useful for economic models that aim to identify the most cost-efficient intervention option and monitor the impact of interventions. We present here the index for goat production in dryland systems in Ethiopia; the approach can easily be adapted to other production systems elsewhere.

Influenza A Virus Infections in Dromedary Camels, Nigeria and Ethiopia, 2015-2017.

We examined nasal swabs and serum samples acquired from dromedary camels in Nigeria and Ethiopia during 2015-2017 for evidence of influenza virus infection. We detected antibodies against influenza A(H1N1) and A(H3N2) viruses and isolated an influenza A(H1N1)pdm09-like virus from a camel in Nigeria. Influenza surveillance in dromedary camels is needed.

Isolation and identification of Brucella melitensis using bacteriological and molecular tools from aborted goats in the Afar region of north-eastern Ethiopia.

BACKGROUND: Infection with Brucella melitensis (B. melitensis) is one of the most important causes of abortion in goats and sheep, and also causes severe systemic disease in exposed humans. In Ethiopia, based on seroepidemiological studies, brucellosis is known to be endemic. However, there is little information on the isolation and molecular detection of Brucella species in small ruminants. Therefore, the present study was conducted in the Amibara district of Afar Region of Ethiopia to isolate and molecularly detect Brucella infection in small ruminants. RESULTS: Out of the total 64 samples cultured, eight samples (five vaginal swabs and three milk) were positive for Brucella species based on colony morphology, growth characteristics, modified acid fast staining and biochemical tests results. Further identification using Brucella- ladder PCR method showed that four of the isolates (three from vaginal swabs and one from milk) from goats amplified fragments of 1071 bp, 794 bp, 587 bp, 450 bp and 152 bp in band size. The molecular result combined with the microbiological and biochemical characteristics of the isolates indicated that the isolates were strains of B. melitensis. CONCLUSION: The finding of this study could suggest economic and zoonotic significance of B. melitensis and warrants for the need for control strategies in livestock and creation of awareness in the pastoral communities on the safe consumption of foods of animal origin and avoidance of physical contact with aborted materials.

Molecular characterization of Mannheimia haemolytica isolates associated with pneumonic cases of sheep in selected areas of Central Ethiopia.

BACKGROUND: Mannheimia haemolytica has been recognized as the principal cause of pneumonic pasteurellosis in sheep and goats. It is one of the important diseases of small ruminants in Ethiopia. While annual vaccination using a monovalent vaccine (inactivated Pasteurella multocida biotype A) is common, respiratory diseases are still reported in various parts of Ethiopia. This suggests the need for further investigation into the species and strains responsible for the disease, which is vital information for development of a multivalent vaccine. The objective of the current study was to isolate M. heamolytica associated with pneumonic cases of sheep in selected areas of Central Ethiopia, determine its role and the strains/genotypes of the bacterium circulating in the study area. RESULTS: Bacteriological analysis of nasal swab samples collected from a total of 76 pneumonic cases of sheep showed that M. haemolytica was isolated from 26 of them while B.trehalosi from two cases. Further molecular analyses of the isolates using M. haemolytica species-specific and M.haemolytica serotype-1 antigen specific PCR assays revealed, 26 of the isolates were identified as M. haemolytica of which 21 of them were M. haemolytica serotype-1. Both M. haemolytica and B.trehalosi isolates were not detected in a PCR assay targeting capsular biosynthesis gene (capA) of P.multocida despite the non-specific products observed in M. haemolytica isolates. Phylogenetic analysis of M. haemolytica isolates included in this study in comparison with the reference strains with respect to PHSSA and Rpt2 genes revealed that the Ethiopian M. haemolytica isolates constituted three distinct genotypes consistent with site of origin. CONCLUSION: The study indicated that M.haemolytica is commonly associated with cases of pneumonia in sheep in the study areas of central Ethiopia although the remaining other pathogens responsible for majority of the cases are yet to be determined. Molecular characterization revealed the existence of three genotypes of M. haemolytica circulating in the study areas consistent to the site of isolation. The findings suggest further extensive work to determine all pathogens associated with sheep pneumonia and the strain distribution of M. heamolytica to understand its molecular epidemiology at national level and design cost effective prevention and control methods.

Appraisal of interpretation criteria for the single intra-dermal comparative cervical tuberculin test for the diagnosis of tuberculosis in dromedary camels in Ethiopia.

Dromedary camels are the main sources of milk, meat and income for the Ethiopian pastoralists as they withstand the harsh environments of the regions of the country. Tuberculosis (TB) affects dromedary camels causing morbidity and mortality in these animals. Hence, early diagnosis and identification of infected camels play a significant role in reducing the transmission of TB in camels. This study was conducted on 168 camels between October 2014 and July 2015 to evaluate the performance of single intra-dermal comparative cervical tuberculin (SICCT) to diagnose TB in camels. Gross pathology was used as a gold standard to define disease status of each camel. The result showed that at the cutoff value of ≥ 3 mm SICCT had optimum performance with sensitivity and specificity of 60.7 and 85%, respectively. Moreover, at a cutoff ≥ 3 mm, the receiver operating characteristics (ROC) revealed area under the ROC curve was 0.729 (0.615-0.842) which is statistically significant (p = 0.000). Thus, the result of the present study could suggest the use of ≥ 3 mm cutoff value for the diagnosis of TB in dromedary camels in Ethiopia.

Molecular typing of Mycobacterium tuberculosis complex isolated from pulmonary tuberculosis patients in central Ethiopia.

BACKGROUND: Identification of the types of strains of Mycobacterium tuberculosis (M. tuberculosis) complex causing tuberculosis (TB) could contribute to TB control program of specific geographic region as well as it could add knowledge onto the existing literature on TB worldwide. The objective of the present study was to identify the species and strains of M. tuberculosis complex causing pulmonary tuberculosis in central Ethiopia. METHODS: A health institution- based cross-sectional study was conducted on 338 smear positive TB cases visiting three hospitals between October 2012 and September 2013. Morning and spot sputum samples were collected before the starting of treatment regimens. Thus, a total of 338 pooled sputum samples collected from these cases. Samples were cultured on Löwenstein Jensen media and the isolates were identified by the region of difference (RD) 9 based polymerase chain reaction (PCR) and spoligotyping. RESULT: Of the total isolates 98.6% of the isolates were identified to be M. tuberculosis while the remaining 1.4% were identified as M. africanum. Further, typing of M. tuberculosis using spoligotyping lead to the identification of 90 different strains of M. tuberculosis. Of these strains, 32 were clustered consisting of more than one isolate while the remaining 58 strains were unique consisting of single isolate. Thus, 79.3% (223/281) of the isolates were found in the clustered while only 20.6% (58/281) of the strains were unique. Forty-five of the spolgotyping patterns were registeredin the SITVIT2 or SpolDB4 database in while the remaining 45 were notfound in the database and hence were orphan strains. The dominant strains were SIT53, SIT149, and SIT54, consisting of 43, 37 and 34 isolates, respectively. Classification of the spoligotype patterns using TB-insight RUN TB-Lineage showed that 86.8, 6.4, 5, 1.4% ofthe isolatesbelonged to the Euro-American lineage, East-African-Indian, Indo-oceanic and M. africanum, respectively. CONCLUSION: The identification of clustered and new strains using spolygotyping in present study does not give conclusive finding as spoligotyping has low discriminatory power. Thus, further identification of these isolates using mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VENTR) and or whole genome sequencing (WGS) recommended.

Nontuberculosis mycobacteria are the major causes of tuberculosis like lesions in cattle slaughtered at Bahir Dar Abattoir, northwestern Ethiopia.

BACKGROUND: The main cause of bovine tuberculosis (bTB) is believed to be Mycobacterium bovis (M. bovis). Nontuberculosis mycobacteria (NTM) are neglected but opportunistic pathogens and obstacles for bTB diagnosis. This study aimed to isolate and characterize the mycobacteria organisms involved in causing TB-like lesions in cattle in northwestern Ethiopia. RESULTS: A total of 2846 carcasses of cattle were inspected for TB lesions. Ninety six tissues (including lymph nodes such as submandibular, retropharyngeal, tonsilar, mediatinal, bronchial and mesenteric, and organs such as lung, liver and kidney) with suspicious TB lesion(s) were collected and cultured on Lowenstein-Jensen medium. Twenty one showed culture growth, of which only 17 were identified containing acid fast bacilli (AFB) by Ziehl-Neelsen staining. Among the 17 AFB isolates 15 generated a polymerase chain reaction product of 1030 bp by gel electrophoresis based on the 16S ribosomal RNA gene amplification. No M. tuberculosis complex species were isolated. Further characterization by Genotype Mycobacterium CM assay showed 6 isolates identified as M. peregrinum. Eight isolates represented by mixed species, which includes M. fortuitum-peregrinum (3 isolates), M. gordonae-peregrinum (3 isolates) and M. fortuitum-gordonae-peregrinum (2 isolates). One NTM could not be interpreted. CONCLUSION: A significant number of NTM species were isolated from TB-like lesions of grazing cattle slaughtered at Bahir Dar Abattoir. Such finding could suggest the role of NTM in causing lesions in cattle. Further investigations are recommended on the pathogenesis of the reported NTM species in cattle, and if they have public health significance.

T cell response to alpha crystallin and Mycobacterium tuberculosis specific antigens using ex-vivo elispot assay for detecting latent tuberculosis infection in Addis Ababa, Ethiopia.

BACKGROUND: Mycobacterium tuberculosis (Mtb) persists in a state of non-replication or stationary phase, but resulting in active tuberculosis (TB) when the immune system is suppressed. Alpha-crystallin (ACR) is one of the bacterial antigens characterized known to be related to shifting of the bacilli from growth to a non-replicating persistent state. OBJECTIVE: To compare the ex-vivo responsiveness of active TB patients, close household contacts and healthy controls to specific Mtb antigens. METHODOLOGY: Antigen-specific interferon-gamma (IFN-g) responses were measured to a 16kDa-alpha crystallin (ACR) antigen along with its peptides and other Mtb antigens (ESAT-6, CFP-10, PPD, TB10.3 and Ag85A) in 39 active TB patients, 23 close household contacts and 25 community controls, using ex-vivo ELISPOT RESULT: The proportion of responders to ACR was 36% in active TB patients (76 +/- 14 spot forming cells), 48% in close household contacts (123 +/- 31 spot forming cells) and 76% in community controls (165 +/- 29 spot forming cells) indicating the presence of latency more in the community controls compared to the other groups. Sixty percent of community controls (131 +/- 27 spot forming cells), 61% of healthy household contacts (138 +/- 3 spot forming cells) and 54% of TB patients (198 +/- 37 spot forming cells) showed ESAT-6-specific T cell responses. CONCLUSION: Antigen specific T cell response based on ex-vivo ELISPOT assay using combined ACR and ESAT-6/ CFP-10 antigens can be used as indicator of underlying latent TB infection in tropical setting where tuberculosis is endemic.

Seroprevalence of Brucellosis, Knowledge, and Risky Practices in Dairy Cattle Owners and Workers in Maekel and Debub Regions, Eritrea.

Brucellosis is a zoonotic disease with worldwide distribution. In Eritrea, the status of the disease in occupationally exposed dairy farmers is unknown. The objective of this study was to determine the seroprevalence of brucellosis, level of knowledge, and risky practices of dairy cattle owners/workers in Maekel and Debub regions, Eritrea. A cross-sectional study was conducted between August 2021 and February 2022. A total of 416 dairy cattle owners and workers underwent blood collection and interview using a standardized questionnaire. Blood samples were tested using Rose Bengal Plate Test, and positive samples were confirmed using competitive ELISA. Variation in knowledge scores by sociodemographic factors and practices were explored statistically. The apparent and true seroprevalence was 1.2% (95% CI: 0.05-2.8%) and 1.4% (95% CI: 0.6-3.4%), respectively. Apparent seroprevalence was similar in Maekel (1.1%) and Debub (1.2%) regions. Nearly half of the participants (49.5%) had never heard of brucellosis before. Overall, brucellosis knowledge score was low (mean score: 6.53/20). Knowledge score was higher in participants from Maekel region (P <0.001), older participants (P = 0.035), those with higher educational attainment (P = 0.001), and those with more years of experience working in dairy farming (P = 0.001). Knowledge score was lower in farm workers compared with family members (P = 0.016). No significant differences in knowledge score existed between participants who engaged in or did not engage in potential risky practices. In summary, the prevalence of brucellosis in dairy cattle owners/workers in Maekel and Debub regions, Eritrea, was low. Participants demonstrated limited knowledge of brucellosis and engaged in risky practices.

Association of the level of IFN-γ produced by T cells in response to Mycobacterium tuberculosis-specific antigens with the size of skin test indurations among individuals with latent tuberculosis in a highly tuberculosis-endemic setting.

There is growing evidence showing the potential of T-cell-based gamma interferon (IFN-γ) release assays (IGRAs) for predicting the risk of progression of Mycobacterium tuberculosis (Mtb) infection, though there is little information from tuberculosis (TB)-endemic settings. In this study, we assessed the association between the level of IFN-γ produced by T cells in response to Mtb-specific antigens and the size of skin test indurations in 505 adult individuals who were screened for latent tuberculosis infection (LTBI) using the QuantiFERON-TB Gold In Tube (QFTGIT) assay and tuberculin skin test (TST). There was a strong positive correlation between the level of IFN-γ induced by the specific antigens and the diameter of the skin indurations (Spearman's rho = 0.6, P < 0.001). Body mass index and parasitic infection were not associated with the level of IFN-γ production or the TST reaction. In linear regression analysis, the size of the skin test indurations was significantly associated with the mean level of IFN-γ [coefficient, 0.65; 95% confidence interval (CI), 0.47 to 0.82, P < 0.001]. Similarly, results from logistic regression analysis demonstrated that individuals who had skin test indurations ≥ 10 mm were 6.82 times more likely than individuals who had skin test indurations < 10 mm to have high levels of IFN-γ (i.e. positive QFTGIT result) (adjusted odd ratio = 6.82; 95% CI, 3.67 to 12.69, P < 0.001). In conclusion, the results of this study could provide indirect evidence for the prognostic use of the QFTGIT assay for progression of Mtb infection, though prospective follow-up studies are needed to provide direct evidence.

Seroprevalence of Bovine Brucellosis in Selected Sites of Central Highland of Ethiopia.

Background: Brucellosis is a contagious, economically significant bacterial disease that affects animals worldwide and is one of the most neglected zoonotic diseases in the world. The disease poses a barrier to the trade of animals and animal products, represents a public health hazard, and is an impediment to free animal movement. Methods: A cross-sectional study was carried out from December 2019 to May 2020 in order to determine seroprevalence and identify potential risk factors for brucellosis in dairy cows in the Central Highlands of Ethiopia with recent cases of abortion. Purposive sampling was carried out on the farms and kebeles in question to screen for recent cases of abortion in dairy cows. For the purpose of performing serological testing, 352 blood samples from dairy cattle were obtained. The Rose Bengal Plate test was used to initially screen the serum samples, and the Complement Fixation test was utilized as a confirmatory test. Results: Using combined RBPT and CFT tests, the overall seroprevalence of bovine brucellosis was 0.6% (95% CI: 0.16-2.09). Retained fetal membrane (OR = 32.74, p = 0.006), market-based stock replacement (OR = 16.55, p = 0.002), breeding method (OR = 7.58, p = 0.027), and late stage of abortion (OR = 14.74, p = 0.0002) are all significantly associated risk factors. Conclusion: The present seroprevalence study revealed that brucellosis is prevalent at a lower rate among dairy cattle in the study areas. However, there is a possible risk of brucellosis transmission in dairy cattle and the exposed human population in research locations because no control measures were put in place there. Implementing a test and slaughter method with compensation for farmers is advised due to the low prevalence of bovine brucellosis in government-owned and small-holder farms.

Seropositivity, Comparison Between the Efficiency of Serological Tests and Risk Factors of Brucella Infection in Small Ruminants with History of Abortion in the Afar Region of North-Eastern Ethiopia.

Purpose: Brucellosis is one of the most important reproductive diseases that cause abortion and breeding failure in small ruminants in Ethiopia. Therefore, our objective was to detect the seropositivity and risk factors of Brucella infection in small ruminants with history of abortion using modified RBPT, cELISA, and CFT in the Amibara district of the Afar Region, Ethiopia. Methods: Sera were collected from 226 animals (195 goats and 31ewes) and assessed for seropositivity of Brucella infection using modified RBPT, CFT, and competitive ELISA. Results: The overall seroprevalence was 12.0% (27 out of 226), 7.5% (17 out of 226), and 26.5% (60 out of 226) by mRBPT, CFT, and cELISA, respectively. Out of 27 sera that were reactive by mRBPT, 17 (63.0%) were also reactive by (CFT). Out of the 17 sera that were reactive by CFT and mRBPT, 14 (82.4%) were reactive by cELISA. Out of the 29 sera that were non-reactive by both mRBPT and CFT, 10 (34.5%) were found reactive by cELISA. Out of the 226 sera that were tested by both mRBPT and cELISA, 20 (8.9%) were reactive by both tests, while 159 (70.4%) were non-reactive by both tests. The percentage of test agreement (79.2%) between mRBPT and cELISA was poor (k=0.353). High seropositivity for Brucella infection was significantly associated with the presence of retained placenta in the studied animals (adjusted OR=2.2, 95% CI, 1.1-4.4, P=0.030) as detected by cELISA. Conclusion: The current study revealed that a cELISA-based seroepidemiological survey increases the likelihood of detecting individuals with brucellosis and provides reliable evidence for mRBPT. Furthermore, there was a significant association between seropositivity for Brucella infection and retained placenta. These findings emphasize the necessity for proactive measures to reduce the economic impact of brucellosis and mitigate the risk of zoonotic transmission.

Comparative Analysis of Tick-Borne Relapsing Fever Spirochaetes from Ethiopia and Nigeria.

Despite increasing reports of tick-borne diseases in Africa, remarkably, reports of tick-borne relapsing fever (TBRF) in Nigeria are lacking. Ornithodoros savignyi from Nigeria have been reported with the relapsing fever Candidatus Borrelia kalaharica. Conversely, in Ethiopia, the agent of relapsing fever is the louse-borne relapsing fever (LBRF) spirochaete Borrelia recurrentis with no TBRF reported to occur. A total of 389 Ornithodoros ticks, Ethiopia (N = 312) and Nigeria (N = 77), were sampled, together with 350 cattle, and 200 goat sera were collected from Nigeria. Samples were screened for Borrelia spp. by RT-PCR. Reactive samples were confirmed, then sequenced using flagellin B, 16S rRNA, and 16S-23S intergenic spacer region. The prevalence of Borrelia spp. in livestock was 3.8% (21/550) and 14% (3/21) after final molecular confirmation. Of 312 ticks from Ethiopia, 3.5% (11/312) were positive for Borrelia, with 36% (4/11) by conventional PCR. Sequencing revealed that the borreliae in soft ticks was C. B. kalaharica, whilst that found in animals was Borrelia theileri. Soft ticks were confirmed by sequencing 7% (22/312) and 12% (9/77) of the Ethiopian and Nigerian ticks, respectively. Phylogenetic analysis revealed that these were Ornithodoros savignyi. This is the first evidence of C. B. kalaharica in Ethiopia and demonstrates the co-existence of TBRF in a country endemic to LBRF. Important, this might cause a diagnostic challenge given that LBRF is predominantly diagnosed by microscopy, which cannot differentiate these two spirochaetes. Furthermore, we report B. theileri in ruminants in Nigeria, which may also be of veterinary and economic importance.

Prevalence of brucellosis and associated risk factors in dairy cattle in Maekel and Debub Regions, Eritrea.

Introduction: Brucellosis is a zoonotic disease with worldwide distribution. It is considered endemic in Eritrea, however, the current prevalence status and related risk factors in animals are unknown. The objective of this study was to determine the prevalence of and risk factors for brucellosis in dairy cattle in Maekel and Debub regions, Eritrea. Methods: A cross sectional study was conducted between August 2021 and February 2022. A total of 2,740 dairy cattle from 214 herds in 10 sub-regions of Eritrea were selected for blood and data collection. Blood samples were tested using Rose Bengal Plate Test (RBPT) and positive samples were confirmed using competitive (c-ELISA). Data on risk factors was collected using questionnaire and analyzed using logistic regression. Results: In total, 34/2740 animals tested positive by RBPT. Of these, 29 were confirmed positive by c-ELISA, giving an apparent and estimated true individual-level prevalence of 1.1% (95% CI: 0.7, 1.5%) and 1.3% (95% CI: 0.9, 1.8%), respectively. Sixteen herds (7.5%) tested positive by RBPT and of these 15 herds (7.0%) were confirmed positive by c-ELISA, giving an estimated true herd-level prevalence of 7.0% (95% CI: 4.0, 10.7). Animal and herd-level apparent prevalence was 1.6 and 9.2% in Maekel, while in Debub it was 0.6 and 5.5%, respectively. Multivariable regression analysis indicated that non-pregnant lactating cows (adjusted odds ratio [aOR] = 3.35; p = 0.042) were more likely to be Brucella sero-positive. History of abortion on the farm (aOR = 5.71; p = 0.026) and larger number of cows in the herd (aOR = 1.14; p < 0.001) were associated with brucellosis sero-positivity in herds. Conclusion: Brucellosis prevalence was low in the study areas. Nonetheless, this low prevalence may increase if the disease is not controlled. Therefore, testing animals before movement, good farming practices, sanitary measures, and an awareness raising program on brucellosis are recommended.