Seroprevalence and associated risk factors of equine herpesvirus type-1/-4 in selected districts of Northwest Amhara, Ethiopia.

This study aimed to estimate the prevalence, determine the distribution, and identify the epidemiological risk factors of EHV-1/-4 infections in selected districts of Northwest Amhara Region. 460 serum samples were collected from equines using multistage cluster sampling technique, and a competitive Enzyme-linked immunosorbent assay (cELISA) was performed. Various risk factors for the occurrence of EHV-1/-4 were considered. Statistical analysis was performed using R version 4.3.1. 65.9% (303) equids were tested positive for antibodies against EHV-1/-4. Based on district, the highest prevalence was recorded in Wogera (86.1%), while the lowest was in Debark (47.4%). There was a significant difference (p <0.05; 95% CI: 1.1067993-3.682843) in the prevalence of EHV-1/-4 among species and donkeys are 2.019 times more likely to get an EHV infection than horses. The prevalence of EHV-1/-4 was highest in equids with the age of 3-8 years and lowest in < 3 years, and the difference was statistically significant (p <0.05; 95% CI: 1.9812042-6.771820). Statistically significant variation (p <0.05; 95% CI: 1.1173822-2.684013) was also observed between sex of equids in which females had 1.73 times higher chance to get EHV infection than males. Higher prevalence was found in lactating equids (81.6%), followed by pregnant equids (74.6%), and dry equids (66.4%). Generally, this study indicated a high and wide distribution of EHV-1/-4 infection in the study area, which needs due attention. Devising strategies to prevent and minimize the spread and occurrence of the infection is crucial.

Strain diversity and gene mutations associated with presumptive multidrug-resistant Mycobacterium tuberculosis complex isolates in Northwest Ethiopia.

OBJECTIVES: In this study, we assessed the genetic diversity and gene mutations that confer resistance to rifampicin (RIF), isoniazid (INH), fluoroquinolone (FQ), and second-line injectable (SLI) drugs in RIF-resistant (RR)/multidrug-resistant tuberculosis (MDR-TB) isolates in Northwest Ethiopia. METHODS: Spoligotyping was used to assign isolates to TB lineages (Ls), and Hain line probe assays were used to detect resistance to RIF, INH, and FQs, and SLIs. RESULTS: Among 130 analyzed strains, 68.5% were RR, and four major Mycobacterium tuberculosis complex lineages (L1, L3, L4, and L7) were identified with a predominance of the Euro-American L4 (72, 54.7%), while L7 genotypes were less common (3, 2.3%). Overall, the L4-T3-ETH (41, 32.0%), L3-CAS1-Delhi (29, 22.7%), and L3-CAS1-Killi (19, 14.8%) families were most common. Line probe analysis showed that among rpoB mutants, 65.2% were S450L, while 87.8% of katG mutants were S315T. Only three isolates showed mutation (c-15t) at the inhA gene, and no double mutation with katG and inhA genes was found. Six strains, two each of L1, L3, and L4, were resistant to FQs, having gyrA mutations (D94G, S91P), of which three isolates had additional resistance to SLI (rrs A1401G or C1402T mutations) including one isolate with low-level kanamycin (KAN) resistance. CONCLUSIONS: This study showed a predominance of L4-T3-ETH, L3-CAS1-Delhi, and L3-CAS1-Killi families, with a high rate of rpoB_S450L and katG_S315T mutations and a low proportion of gyrA and rrs mutations. L7 was less frequently observed in this study. Further investigations are, therefore, needed to understand L7 and other lineages with undefined mutations.

Isolation and Molecular Detection of Pasteurellosis from Pneumonic Sheep in Selected Areas of Amhara Region, Ethiopia: An Implication for Designing Effective Ovine Pasteurellosis Vaccine.

Introduction: Pneumonic pasteurellosis mainly caused by bacterial species of Mannheimia, Pasteurella, and Bibersteinia causes a significant financial loss to the sheep production sector through reduced productivity and high mortality. There is a dearth of information on the major agents involved in the disease in the Amhara region, Ethiopia. Therefore, the aim of this study was to isolate and molecularly confirm Mannheimia, Pasteurella, and Bibersteinia from nasal swabs of sheep suspected of pneumonic pasteurellosis in selected areas of the Amhara region. Methods: Isolation and phenotypic characterization were performed using microbiological and biochemical testing according to standard methods. Molecular confirmation of isolates was done through amplification of virulence associated genes, PHSAA and Rpt2, of Mannheimia hemolyticausing multiplex PCR. Results: Accordingly, 46 out of 141 (32.62%) samples were presumably identified as M. hemolytica with no Pasteurella multocida and Bibersteinia trehalosi. Seven (n=7) out of the 46 isolates tested positive for either of the two virulence genes. Discussion and conclusion: The finding of this study is indicative that M. hemolytica is the main bacteria linked with pneumonic pasteurellosis in the study area which suggests the need to develop a polyvalent vaccine including strains of M. hemolytica or its antigenic determinants. However, the role of other bacterial, viral, and parasitic agents in the cases investigated should also be considered.

Prevalence of bovine tuberculosis and distribution of tuberculous lesions in cattle slaughtered at Gondar, Northwest Ethiopia.

BACKGROUND: In Ethiopia, bovine tuberculosis (BTB) is a neglected disease that affects the economy and livelihoods of farmers. However, the available data is limited due to insufficient disease surveillance in the country. Therefore; this study aimed to assess the prevalence and distribution of lesions of BTB in cattle slaughtered at Gondar, Northwest Ethiopia. METHODS: Postmortem examinations were used to detect tuberculous lesions, while smear microscopy and histopathology were performed for the identification of acid-fast bacilli (AFB). RESULTS: Of 497 inspected slaughtered cattle, 45 (9.1%, 95%CI; 0.0668-0.1193) were diagnosed with BTB suggestive tuberculous lesions. A higher proportion of gross lesions was recorded in lymph nodes of lungs; at the mediastinal (14, 31.1%) and bronchial (10, 22.2%) lymph nodes, and followed by mesenteric lymph nodes (9, 20%). Of 45 tuberculous lesions; only 2 (4.4%) were identified as AFB positive by smear microscopy and histopathology. In the overall statistical analysis, body conditions of slaughtered cattle were found to be significantly associated with BTB tuberculous lesions (p < 0.05). CONCLUSION: This finding provides the prevalence of BTB and distribution of tuberculous lesions in cattle slaughtered at the abattoir and highlights the need for a practicable control strategy of the disease in the region.

Bacteriological and molecular characterization of Mycobacterium bovis isolates from tuberculous lesions collected among slaughtered cattle, Northwest Ethiopia.

BACKGROUND: In Ethiopia, the distribution of bovine tuberculosis (BTB) has long been known and documented as a major problem of animal health. However, the burden of circulating M. bovis strains is poorly understood in the country. Therefore; this study aimed to identify and characterize the mycobacterial isolates responsible for BTB in Northwest Ethiopia. METHODS: A cross-sectional study was conducted on tuberculous lesions that had been collected from slaughtered cattle between September 2018 to June 2019. Collected lesions were cultured and tested for tuberculous bacilli. The MPT64 assay and Genotype line probe assay (LPA) were used for identification of mycobacterial isolates, and region of deletion 4 (RD4) typing and spoligotyping were used to characterize the M. bovis strains. RESULTS: Of the total 1458 examined slaughtered cattle, only 62 (4.3, 95%CI; 0.0328-0.0542) had tuberculous lesions. The highest number of gross tuberculous lesions were observed from the lymph nodes of the thoracic cavity; at the mediastinal (40.3%, 25/62) and bronchial (22.6%, 14/62) lymph nodes. Of the 62 collected tuberculous lesions; 18 (29.0%) were culture positive for mycobacterium isolates, and only five isolates were confirmed for M. tuberculosis complex (MTBc) by the MPT64 assay and LPA. All the five MTBc isolates were positive for RD4 typing of M. bovis with a PCR product size of 446 bp, and no isolate was noticed to have M. tuberculosis. The detected M. bovis strains displayed five spoligotypes; with the common SB1176 and SB0133 M. bovis strains, although the two spoligotypes had not been previously reported. CONCLUSION: The present study shows that BTB in North Gondar, Ethiopia, is caused by M. bovis strains SB1176 and SB0033, with low frequency. Thus, the finding highlights the importance of continuous surveillance for mycobacterial strains in cattle populations.

Prevalence, Isolation, Identification, and Risk Factors of Major Bacterial Cause of Camel Subclinical Mastitis.

OBJECTIVE: A crosssectional study was conducted between September 2015 and August 2016 in the district of Afar Regional State, Northeastern Ethiopia, to characterize the most prevalent bacterial pathogens and identify the associated risk factors of camel subclinical mastitis. California mastitis test (CMT) was used as a screening test, and standard bacteriological methods were carried out for isolation and identification of the pathogens. RESULTS: Among the total 96 lactating camels examined, 25 were found positive with the overall prevalence of 26%, with 25% and 1% subclinical and clinical mastitis cases, respectively. Totally, 384 quarters of udder were examined; of these, 10 of them were blind while the rest 374 were nonblind teats. The quarter level prevalence of subclinical mastitis was 8.9%. The analysis showed that statistically significant difference (P < 0.05) of tick infestation and subclinical mastitis. Additionally, among the bacteriologically tested 34 CMT positive milk samples, all of them showed growth on nutrient and blood agar plate. Out of these culture isolates, the major bacterial pathogens identified were Staphylococcus aureus (8.7%), Staphylococcus hyicus (6.52%), Staphylococcus intermedius (6.52), Coagulase-negative staphylococci (19.57%), Bacillus (19.57%), Escherichia coli (6.52%), and Pasteurella multocida (6.52%) species. Therefore, appropriate control measures and awareness creation to the community should be practiced.

Genetic diversity of the Mycobacterium tuberculosis complex strains from newly diagnosed tuberculosis patients in Northwest Ethiopia reveals a predominance of East-African-Indian and Euro-American lineages.

OBJECTIVES: This study described the population structure of M. tuberculosis complex (MTBc) strains among patients with pulmonary or lymph node tuberculosis (TB) in Northwest Ethiopia and tested the performance of culture isolation and MPT64-based speciation for Lineage 7 (L7). METHODS: Patients were recruited between April 2017 and June 2019 in North Gondar, Ethiopia. The MPT64 assay was used to confirm MTBc, and spoligotyping was used to characterize mycobacterial lineages. Line probe assay (LPA) was used to detect resistance to rifampicin and isoniazid. RESULTS: Among 274 MTBc genotyped isolates, there were five MTBc lineages: L1-L4 and L7 were identified, with predominant East-African-Indian (L3) (53.6%) and Euro-American (L4) (40.1%) strains, and low prevalence (2.6%) of Ethiopia L7. The genotypes were similarly distributed between pulmonary and lymph node TB, and all lineages were equally isolated by culture and recognized as MTBc by the MPT64 assay. Additionally, LPA showed that 259 (94.5%) MTBc were susceptible to both rifampicin and isoniazid, and one (0.4%) was multi-drug resistant (resistant to both rifampicin and isoniazid). CONCLUSION: These findings show that TB in North Gondar, Ethiopia, is mainly caused by L3 and L4 strains, with low rates of L7, confirmed as MTBc by MPT64 assay and with limited resistance to rifampicin and isoniazid.

In Vitro Anthelmintic Activity of Crude Extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus.

Gastrointestinal nematodes (GINs) are the major limiting factor for the successfulness of livestock production throughout the world. Emergence of resistance strains as well as scarcity and high cost of the currently available drugs has led to the evaluation of other alternative helminth control options, mainly from plants. The current study is aimed at investigating the in vitro anthelmintic efficacy of crude methanolic extracts of two traditionally important medicinal plants, Artemisia herba-alba and Punica granatum, against Haemonchus contortus using adult motility assay (AMA) and egg hatch inhibition assay (EHIA). Four graded concentrations of the extracts were tested for both the AMA (10, 5, 2.5, and 1.25 mg/mg) and EHIA (0.1, 0.25, 0.5, and 1 mg/mL) in replicates. Albendazole and phosphate-buffered saline (AMA) or distilled water (EHIA) were used as the positive and negative controls, respectively. The crude extracts of A. herba-alba and P. granatum exhibited a potential anthelmintic activity at all dose levels in a concentration- and time-dependent fashion. The highest concentration (10 mg/mL) of all the extracts caused a significantly (p < 0.05) superior nematocidal activity compared to the negative control. Moreover, significant and concentration-dependent egg hatching inhibition effect was observed from both plant extracts. Maximal (98.67%) egg hatching inhibition effect was exhibited by the flower extract of A. herba-alba at 1 mg/mL concentration. The relative egg hatch inhibition efficacy indicated that both plants caused a significantly (p < 0.05) greater egg hatch inhibition within 48 hr of exposure. The current study validated the traditional use of both plants as a natural anthelmintic against H. contortus justifying a need to undertake detail pharmacological and toxicological investigation on both plants.

Multidrug-resistant patients receiving treatment in Niger who are infected with M. tuberculosis Cameroon family convert faster in smear and culture than those with M. tuberculosis Ghana family.

In this study, we analyzed the M. tuberculosis complex (MTBc) population structure among multidrug-resistant TB (MDR-TB) patients in Niger and tested whether the Cameroon family displayed a slower response to MDR-TB treatment. We genotyped baseline clinical isolates that had been collected from pulmonary MDR-TB patients recruited consecutively between 2008 and 2016 in Niger. Spoligotyping was used to analyze the genetic diversity of mycobacterial lineages, and Kaplan Meier's analysis to compare treatment outcomes. A total of 222 MTBc isolates were genotyped; 204 (91,9%) were identified as the Euro-American L4 lineage, with the Ghana family (106, 47,4%) and the Cameroon family (63, 28,4%) being predominant. Patients infected by Cameroon family isolates 61(96,8%) showed faster conversion (log-rank p < 0.01) than those infected with Ghana family isolates (91,5%), and were more likely to experience favorable outcome (adjusted odds ratio [aOR] 4.4; 95%CI 1.1-17.9]; p = 0.015). We found no association between MTBc families and second-line drug resistance profiles (p > 0.05). Our findings show that MDR-TB in Niger is caused by major spoligotypes of the Euro-American L4; with more rapid smear and culture conversion in patients infected with the Cameroon family. These first insights may alert clinicians that slow conversion may be associated with the type of infecting strain.

Seroprevalence and Risk Factors Associated with Contagious Caprine Pleuropneumonia in Western Amhara, Northwest Ethiopia.

Contagious caprine pleuropneumonia (CCPP) has been identified as a significant problem in goat production, especially in the arid and semiarid lowland areas of Ethiopia. Even though CCPP was reported in most of the goat rearing areas of the country, there is no adequate information on the disease in the Amhara Region. Cross-sectional study was conducted from November 2016 to April 2017 in the districts of Western Amhara to estimate the seroprevalence and identify the associated risk factors for occurrence of the CCPP. The risk factors considered included age, sex, agroclimate, and districts. A competitive enzyme-linked immunosorbent assay (cELISA) was carried out on a total of 400 goat sera samples, out of which 34 samples were found seropositive for specific antibodies against CCPP, with the overall seroprevalence of 8.5% (95% confidence interval (CI) =5.8, 11.2). Among the epidemiological factors considered, age and sex of the goats were not significantly associated with CCPP seroprevalence (p>0.05). However, the seropositivity was slightly higher in adults (9.9%) and female goats (9.0%) compared to young (6.3%) and male (7.5%) goats, respectively. The analysis of seroprevalence by district shows that the seroprevalence of CCPP in Metema (OR=14.34; 95%CI= 1.80, 114.09; p=0.012) and Fogera (OR=9.99; 95%CI= 1.10, 91.16; p= 0.041) was significantly higher compared to other study districts. Multivariable logistic regression analysis also identified the district as a risk factor for the occurrence of a high seroprevalence of CCPP. The present study revealed the seroprevalence and the distribution of CCPP in Western Amhara districts, and hence appropriate control measures including regular investigation and vaccination should be implemented to alleviate the problem.

A Mycobacterial Perspective on Tuberculosis in West Africa: Significant Geographical Variation of M. africanum and Other M. tuberculosis Complex Lineages.

BACKGROUND: Phylogenetically distinct Mycobacterium tuberculosis lineages differ in their phenotypes and pathogenicity. Consequently, understanding mycobacterial population structures phylogeographically is essential for design, interpretation and generalizability of clinical trials. Comprehensive efforts are lacking to date to establish the West African mycobacterial population structure on a sub-continental scale, which has diagnostic implications and can inform the design of clinical TB trials. METHODOLOGY/PRINCIPAL FINDINGS: We collated novel and published genotyping (spoligotyping) data and classified spoligotypes into mycobacterial lineages/families using TBLineage and Spotclust, followed by phylogeographic analyses using statistics (logistic regression) and lineage axis plot analysis in GenGIS, in which a phylogenetic tree constructed in MIRU-VNTRplus was analysed. Combining spoligotyping data from 16 previously published studies with novel data from The Gambia, we obtained a total of 3580 isolates from 12 countries and identified 6 lineages comprising 32 families. By using stringent analytical tools we demonstrate for the first time a significant phylogeographic separation between western and eastern West Africa not only of the two M. africanum (West Africa 1 and 2) but also of several major M. tuberculosis sensu stricto families, such as LAM10 and Haarlem 3. Moreover, in a longitudinal logistic regression analysis for grouped data we showed that M. africanum West Africa 2 remains a persistent health concern. CONCLUSIONS/SIGNIFICANCE: Because of the geographical divide of the mycobacterial populations in West Africa, individual research findings from one country cannot be generalized across the whole region. The unequal geographical family distribution should be considered in placement and design of future clinical trials in West Africa.

Prevalence and Antimicrobial Resistance of Salmonella Isolated from Animal-Origin Food Items in Gondar, Ethiopia.

Salmonella has been found to be the major cause of foodborne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. A cross-sectional study was conducted between February 2014 and December 2015 on food items of animal origin to assess the prevalence and antimicrobial resistance profiles of Salmonella isolates using standard bacteriological methods. The overall prevalence rate of 5.5% was recorded from the total analyzed food items of animal origin. Salmonella isolates were detected from 12% of raw meat, 8% of minced meat, 2.9% of burger samples, 18% of raw eggs, and 6% of raw milk. Furthermore, antimicrobial susceptibility test identified 47.6% resistant Salmonella isolates, 28.6% intermediately sensitive isolates, and 23.8% susceptible isolates. Among Salmonella isolates tested, 42.6%, 28.6%, and 14.3% were found to be relatively resistant to tetracycline, sulfamethoxazole-trimethoprim, and ampicillin, respectively, while 9.5%-19% were intermediately resistant to tetracycline, amoxicillin, ampicillin, cephalothin, and nitrofurantoin. Therefore, our findings provide the prevalence and drug resistance of Salmonella from foods of animal origin and contribute information to scientists as well as public health researchers to minimize the prevalent and resistant foodborne Salmonella species in Ethiopia.

Prevalence and antimicrobial susceptibility profile of listeria species from ready-to-eat foods of animal origin in Gondar Town, Ethiopia.

BACKGROUND: Listeriosis, mostly caused by Listeria monocytogenes species, has become a major concern to public health authorities due to its clinical severity and high mortality rate, particularly in high risk groups. Currently, there is limited information regarding the prevalence and antimicrobial susceptibility profiles of listeria species in ready-to-eat foods of animal origin in Gondar town, Ethiopia. The aim of this study was to determine the prevalence and antimicrobial susceptibility pattern of Listeria species isolated from ready-to-eat food of animal origin from public dinning places in Gondar town, Ethiopia. A cross sectional study on ready-toeat foods of animal origin sampled from major supermarkets, butcher shops, pastry shops, restaurants and hotels was carried out. Culture, biochemical and sugar tests were conducted for listeria species identification and disc diffusion test was performed to study the antimicrobial susceptibility profiles of the isolates. RESULTS: Out of 384 food samples examined, 96 (25%) were positive for Listeria species. Listeria monocytogenes was detected in 24 (6.25%) of the samples. Listeria monocytogenes was isolated from cake, raw meat, ice cream, minced beef, fish, unpasteurized milk and pizza in that order from higher to lower rate. Assessment of antimicrobial susceptibility profile of L. monocytogenes revealed the presence of four multi-drug resistant isolates. The higher resistance rate was recorded for penicillin, nalidixic acid, tetracycline and chloramphenicol, in decreasing order. All L. monocytogenes identified in the current study were sensitive to amoxicillin, cephalothin, cloxacillin, sulfamethoxazole, gentamicin and vancomycin. CONCLUSIONS: The presence of L. monocytogenes including drug resistant and multidrug resistant isolates in some ready-to-eat food items is an indicator of the presence of public health hazards to the consumer, particularly to the high-risk groups. Hence awareness creation on food safety and implementation of regulations about the use of drugs in humans and animals is strongly recommended.

First insights into circulating Mycobacterium tuberculosis complex lineages and drug resistance in Guinea.

In this study we assessed first-line anti-tuberculosis drug resistance and the genotypic distribution of Mycobacterium tuberculosis complex (MTBC) isolates that had been collected from consecutive new tuberculosis patients enrolled in two clinical trials conducted in Guinea between 2005 and 2010. Among the total 359 MTBC strains that were analyzed in this study, 22.8% were resistant to at least one of the first line anti-tuberculosis drugs, including 2.5% multidrug resistance and 17.5% isoniazid resistance, with or without other drugs. In addition, further characterization of isolates from a subset of the two trials (n = 184) revealed a total of 80 different spoligotype patterns, 29 "orphan" and 51 shared patterns. We identified the six major MTBC lineages of human relevance, with predominance of the Euro-American lineage. In total, 132 (71.7%) of the strains were genotypically clustered, and further analysis (using the DESTUS model) suggesting significantly faster spread of LAM10_CAM family (p = 0.00016). In conclusion, our findings provide a first insight into drug resistance and the population structure of the MTBC in Guinea, with relevance for public health scientists in tuberculosis control programs.

Shifts in Mycobacterial Populations and Emerging Drug-Resistance in West and Central Africa.

In this study, we retrospectively analysed a total of 605 clinical isolates from six West or Central African countries (Benin, Cameroon, Central African Republic, Guinea-Conakry, Niger and Senegal). Besides spoligotyping to assign isolates to ancient and modern mycobacterial lineages, we conducted phenotypic drug-susceptibility-testing for each isolate for the four first-line drugs. We showed that phylogenetically modern Mycobacterium tuberculosis strains are more likely associated with drug resistance than ancient strains and predict that the currently ongoing replacement of the endemic ancient by a modern mycobacterial population in West/Central Africa might result in increased drug resistance in the sub-region.