Isolation and characterization of lytic bacteriophages from various sources in Addis Ababa against antimicrobial-resistant diarrheagenic Escherichia coli strains and evaluation of their therapeutic potential.

BACKGROUND: Escherichia coli is a common fecal coliform, facultative aerobic, gram-negative bacterium. Pathogenic strains of such microbes have evolved to cause diarrhea, urinary tract infections, and septicemias. The emergence of antibiotic resistance urged the identification of an alternative strategy. The use of lytic bacteriophages against the control of pathogenic E. coli in clinics and different environmental setups (waste and drink water management) has become an alternative therapy to antibiotic therapy. Thus, this study aimed to isolate and characterize lytic bacteriophage from various sources in Addis Ababa, tested them against antimicrobial-resistant diarrheagenic E. coli strains and evaluated their therapeutic potential under in vitro conditions. METHODS: A total of 14 samples were processed against six different diarrheagenic E. coli strains. The conventional culture and plaque analysis agar overlay method was used to recover lytic bacteriophage isolates. The phage isolates were characterized to determine their lytic effect, growth characteristics, host range activity, and stability under different temperature and pH conditions. Phage isolates were identified by scanning electron microscope (SEM), and molecular techniques (PCR). RESULTS: In total, 17 phages were recovered from 84 tested plates. Of the 17 phage isolates, 11 (65%) were Myoviridae-like phages, and 6 (35%) phage isolates were Podoviridae and Siphoviridae by morphology and PCR identification. Based on the host range test, growth characteristics, and stability test 7 potent phages were selected. These phages demonstrated better growth characteristics, including short latent periods, highest burst sizes, and wider host ranges, as well as thermal stability and the ability to survive in a wide range of pH levels. CONCLUSIONS: The promising effect of the phages isolated in this study against AMR pathogenic E. coli has raised the possibility of their use in the future treatment of E. coli infections.

Genetic Diversity and Population Structure of Selected Ethiopian Indigenous Cattle Breeds Using Microsatellite Markers.

Background: In Ethiopia, livestock contributes 45% of agricultural GDP. Despite the economic role played by the sector, there have been little efforts to genetically improve the indigenous cattle. Morphological characterization of selected Ethiopian indigenous cattle has been made for (Bonga, Jimma, and Kerayu) cattle types. But, the selected indigenous cattle were not characterized at molecular level (genetic diversity information). Hence, this work was initiated to detect and determine the genetic diversity and population structure of selected Ethiopian indigenous cattle ecotypes using microsatellite markers. Results: Different alleles were identified (131) and thirty-three of these alleles were unique to specific ecotypes. All loci used were informative with PIC values ranging from 0.5 (TGLA126) to 0.84 (ETH10) with a mean of 0.70 per locus. The Shannon information index ranged from (I = 1.02) ILST006 to (I = 1.63) ETH10 with an average of 1.28 revealing there is genetic diversity. Moreover, analysis of molecular variance (AMOVA) revealed 84% genetic variation within a population and 13% variation among populations. The value of F-statistics (Fst) (0.129 = 13%) indicated that there was moderate genetic differentiation among ecotypes. The (UPGMA) revealed, Bonga and Jimma clustered together while Kerayu cattle were relatively distinct, Principal coordinates analysis (PCOA) and structure analysis grouped the individual into different clusters confirming the presence of ecotype admixture due to geographical origins and uncontrolled mating. Conclusion: In general, this study has successfully characterized the genetic diversity and population structure of Bonga, Jimma, and Kerayu cattle ecotypes using high polymorphic/informative microsatellite markers. According to this study, Kerayu cattle have high AR and PA when compared to Bonga and Jimma cattle populations. So, the Kerayu population is more diverse than others and it is the hotspot for genetic diversity study. The generated information is very relevant for breeder and genetic conservation.

Treatment of uncomplicated Plasmodium vivax with chloroquine plus radical cure with primaquine without G6PDd testing is safe in Arba Minch, Ethiopia: assessment of clinical and parasitological response.

BACKGROUND: Ethiopia rolled out primaquine nationwide in 2018 for radical cure along with chloroquine for the treatment of uncomplicated Plasmodium vivax malaria in its bid for malaria elimination by 2030. The emergence of anti-malarial drug resistance would challenge the elimination goal. There is limited evidence on the emergence of chloroquine drug resistance. The clinical and parasitological outcomes of treatment of P. vivax with chloroquine plus radical cure using low dose 14 days primaquine were assessed in an endemic area of Ethiopia. METHODS: A semi-directly observed 42-days follow up in-vivo therapeutic efficacy study was conducted from October 2019 to February 2020. Plasmodium vivax mono-species infected patients (n = 102) treated with a 14 days low dose (0.25 mg/kg body weight per day) primaquine plus chloroquine (a total dose of 25 mg base/kg for 3 days) were followed for 42 days to examine clinical and parasitological outcomes. Samples collected at recruitment and days of recurrence were examined by 18 S based nested polymerase chain reaction (nPCR) and Pvmsp3α nPCR-restriction fragment length polymorphism. Asexual parasitaemia and the presence of gametocytes were assessed on the scheduled days using microscopy. Clinical symptoms, haemoglobin levels, and Hillmen urine test were also assessed. RESULTS: Of the 102 patients followed in this study, no early clinical and parasitological failure was observed. All patients had adequate clinical and parasitological responses within the 28 days of follow up. Late clinical (n = 3) and parasitological (n = 6) failures were observed only after day 28. The cumulative incidence of failure was 10.9% (95% confidence interval, 5.8-19.9%) on day 42. Among the paired recurrent samples, identical clones were detected only in two samples on day 0 and day of recurrences (day 30 and 42) using Pvmsp3α genotyping. No adverse effect was detected related to the low dose 14 days primaquine administrations. CONCLUSION: Co-administration of CQ with PQ in the study area is well tolerated and there was no recurrence of P. vivax before 28 days of follow up. Interpretation of CQ plus PQ efficacy should be done with caution especially when the recurrent parasitaemia occurs after day 28. Therapeutic efficacy studies with appropriate design might be informative to rule out chloroquine or primaquine drug resistance and/or metabolism in the study area.

Prevalence of Campylobacter species and associated risk factors for contamination of dairy products collected in a dry season from major milk sheds in Ethiopia.

A cross-sectional study was conducted to investigate the prevalence and risk factors for contamination of Ethiopian dairy products with Campylobacter. A total of 912 dairy food samples were collected from establishments of 682 study participants that were interviewed. Samples were tested for Campylobacter by following the ISO 10272-1:2017 standard and PCR confirmation. Campylobacter was detected in 11% of tested food samples and all detected Campylobacter were C. jejuni. The highest prevalence of C. jejuni was found in raw milk (16%), followed by pasteurized milk (9%) and cottage cheese (2%) (P < 0.001). Using warm water and soap for cleaning cow udders and teats on farms reduced the likelihood of detecting Campylobacter in milk (AOR = 0.3, P = 0.023). Filtering milk with a cloth, using a plastic filter (AOR = 0.065, P = 0.005), and storing milk in an aluminum container (AOR = 0.23, P = 0.027) reduced the likelihood of detecting Campylobacter in milk at the collection facilities. In contrast, Campylobacter detection was significantly more likely in milk collected at collection centers with concrete floors (AOR = 5.2, P = 0.004). The odds of detecting Campylobacter in milk were 17 times greater (AOR = 17, P = 0.007) in milk processing facilities that did not calibrate a pasteurizer on an annual basis. Finally, having a separate refrigerator for milk storage reduced the odds of detecting Campylobacter in retail (AOR = 0.29, P = 0.021).

Aerobic bacteriological studies on the respiratory tracts of apparently healthy and pneumonic camels (Camelus dromedaries) in selected districts of Afar Region, Ethiopia.

A cross-sectional study was conducted to isolate and identify bacterial species from the respiratory tract of apparently healthy and pneumonic camels in Asayita and Dubti woredas in the Afar Region, Ethiopia. From a total of 74 lung tissue and 74 tracheal swab samples Staphylococcus aureus, 16.3%, Streptococcus equi subsp. equi, 13.0%, and Pasteurella multocida, 10.9%, were dominant isolates from pneumonic lungs; Escherichia coli, 12.7%, Proteus species, 10.9%, and Klebsiella pneumoniae, 9.1%, were the majority in the normal lungs. The majority of the isolates colonized both anatomical sites investigated. There was a statistically significant association between the health status of the camels as well as the anatomical site studied with the isolation rates of the major respiratory pathogens (p < 0.05). Furthermore, the isolates were susceptible to norfloxacin, streptomycin, and gentamicin but resistant to ampicillin and tetracycline on in vitro test. Further studies on the pathogenicity of the major isolates are recommended.

Replication characteristics of equine herpesvirus 1 and equine herpesvirus 3: comparative analysis using ex vivo tissue cultures.

Replication kinetics and invasion characteristics of equine herpesvirus-1 and -3 (EHV-1/-3) in nasal and vaginal mucosae were compared using explants. The explants were cultured during 96 h with little change in viability. The tissues were inoculated with EHV-1 03P37 (neuropathogenic), 97P70 (abortigenic) and EHV-3 04P57, collected at 0, 24, 48 and 72 h post inoculation (pi) and stained for viral antigens. Both EHV-1 and EHV-3 replicated in a plaquewise manner. The plaques were already observed at 24 h pi, their size increased over time and did not directly cross the basement membrane (BM). However, EHV-1 infected the monocytic cells (MC) and hijacked these cells to invade the lamina propria. In contrast, EHV-3 replication was fully restricted to epithelial cells; the virus did not breach the BM via a direct cell-to-cell spread nor used infected MC. EHV-1-induced plaques were larger in nasal mucosa compared to vaginal mucosa. The opposite was found for EHV-3-induced plaques. Both EHV-1 strains replicated with comparable kinetics in nasal mucosa. However, the extent of replication of the abortigenic strain in vaginal mucosa was significantly higher than that of the neuropathogenic strain. Two-to-five-fold lower numbers of EHV-1-infected MC underneath the BM were found in vaginal mucosa than in nasal mucosa. Our study has shown that (i) EHV-1 has developed in evolution a predisposition for respiratory mucosa and EHV-3 for vaginal mucosa, (ii) abortigenic EHV-1 replicates better in vaginal mucosa than neuropathogenic EHV-1 and (iii) EHV-3 demonstrated a strict epithelial tropism whereas EHV-1 in addition hijacked MC to invade the lamina propria.

Serological survey of viral diseases relating to reproductive failure among Artiodactyla in Ethiopian Camelus dromedarius.

Serological surveys were performed on Ethiopian camels with a history of abortion to investigate the presence of antibodies against viruses that infect animals classified in the order Artiodactyla. In 2013, 120 serum samples were collected from camels in various parts of Ethiopia. Several viruses related to abortion in ruminants were prevalent. In particular, antibodies against bluetongue virus, were detected at a high rate (76.7% of samples). Additionally, antibodies against Akabane virus and Japanese encephalitis virus were also detected in samples from more than 40% of the camels; however, their antibody titers were relatively low.

Characterization of Escherichia coli isolated from calf diarrhea in and around Kombolcha, South Wollo, Amhara Region, Ethiopia.

This study was carried out from October 2012 to end of February 2013 in and around Kombolcha, Amhara regional state, Ethiopia, using a total of 201 neonatal calves aged 1 day to 4 months and suffering from diarrhea. The objectives of the study were to isolate Escherichia coli from diarrheic calves, and to determine E. coli biotypes and risk factors associated with its isolation. The fecal samples were collected, transported, and processed following standard microbiological procedures. Seventy-four isolates of E. coli were identified. Yellowish diarrhea, younger age, and low-colostrum feeding were significantly associated with rate of E. coli isolation (P < 0.05). Then the 74 isolates of E. coli were biotyped using fermentation of 9 sugars and grouped into 12 biotypes; the most dominant was biotype III (36.8 %). Finally, by comparing with studies elsewhere, from the 12 isolated biotypes, 3 of them were suggested to be enteropathogenic E. coli (EPEC), entherotoxigenic E. coli (ETEC), and adhesion and effacing E. coli (AEEC) pathogenic strains. The present study showed that E. coli accounted for 37 % of calf diarrhea, with very diverse biotypes.

First report of Toxoplasma gondii in camels (Camelus dromedarius) in Ethiopia: bioassay and seroepidemiological investigation.

BACKGROUND: Toxoplasmosis is a major public health concern in many countries of the world. A cross-sectional and follow up experimental study designs were used for seroepidemiological and bioassay studies, respectively from November 2012 to April 2013. The objectives were to estimate the seroprevalence of T. gondii infection, to assess risk factors and to isolate the parasite from camels in the Fentale district, Ethiopia. A direct agglutination test (DAT) and indirect enzyme linked immunosorbent assay (ELISA) kits were used to test camel sera. Hearts and tongues (each 25 g) from 31 seropositive camels were bioassayed in mice. Associations between seroprevalence and potential risk factors (collected using a questionnaire survey) were analyzed using logistic regression. RESULTS: An overall T. gondii prevalence of 49.62% (220/455) by DAT and 40.49% (179/451) by indirect ELISA test were detected. Herd level seroprevalence of 96.77% (30/31) (95% CI: 83.30- 99.92) by DAT was recorded and it was significantly higher in areas where wild felids are present (P = 0.038). Multivariable logistic regression showed that the likelihood of acquiring T. gondii infection was significantly higher in camels in the Ilala pastoral association [PA] (82.26%) (Adjusted Odds ratio [aOR] = 10.8; P < 0.001) than camels in the Galcha PA (31.43%), in camels of ≥ 8 years old (56.52%; aOR = 1.88; P = 0,033) than camels of ≤ 4 years old (34.26%) and in areas where domestic cats are present (aOR = 4.16; P = 0.006). All camel owners were uneducated, handle aborted fetus with bare hands, and drink raw camel milk. DAT and ELISA tests had moderate agreement (Kappa = 0.41). Viable T. gondii were isolated from 16.13% (5/31) of DAT positive camels. One DAT positive but ELISA negative camel sample gave a cyst positive result. CONCLUSIONS: T. gondii infection of camels in the study district is widespread. Age, presence of domestic cats and study PA are independent predictors of T. gondii seropositivity. Isolation of viable parasites from edible tissues of camels and the very poor knowledge of pastoralists about toxoplasmosis suggest the need for prevention of toxoplasmosis through bio-security measures, education and further investigation to unravel the impact of camel toxoplasmosis deserves consideration.

Prevalence of Helicobacter pylori infection and effectiveness of first-line triple eradication therapy among dyspeptic patients at hospitals in Hawassa City, Ethiopia: a cross-sectional follow-up study.

BACKGROUND: Dyspepsia is a common gastrointestinal illness sometimes associated with Helicobacter pylori (H. pylori) infection. Screening and eradicating the bacterium reduces the risk of infection-related complications. The aim of this study was to determine the magnitude of H. pylori infection among dyspeptic patients and the effectiveness of triple eradication therapy at hospitals in Hawassa city, Ethiopia. RESULTS: The prevalence of H. pylori infection was 48.5%. The H. pylori eradication rate using first-line triple therapy was 83.8%. Eradication therapy failure is associated with previous exposure compared to no exposure (AOR: 4.8, 95% CI: 1.37-10.97), a regimen for 10-days compared to 14-days (AOR: 4.05, 95% CI: 1.42-11.55), and self-reported side effects compared to no report (AOR: 2.5, 95% CI: 1.12-5.97). Based on Morisky-eight scale 230 (79.0%) patients were adherent to their triple therapy. Participants with no reports of adverse effects showed increased odds of adherence to triple therapy compared to those who had reports (AOR = 2.45, 95% CI: 1.29-4.62). CONCLUSIONS: This study demonstrated that about half of adult dyspeptic patients were infected with H. pylori, and moderate eradication was observed. Factors such as previous history of eradication therapy, duration of the eradication regimen, and perception of potential adverse effects are associated with eradication rate and should be considered during the initiation of eradication therapy.

Seasonal variation of Salmonella enterica prevalence in milk and cottage cheese along the dairy value chain in three regions of Ethiopia.

Seasonal fluctuations influence foodborne illness transmission and affect patterns of microbial contamination of food. Previous investigations on the seasonality of Salmonella enterica prevalence in dairy products in Ethiopia have been minimal. However, such data are needed to inform strategic development of effective interventions to improve food safety, as seasonal differences may affect intervention strategies. This study was conducted to identify differences in the prevalence of Salmonella in milk and cheese samples between wet and dry seasons. A longitudinal study design was utilized with a random sampling occurring during both dry and wet seasons. A total of 448 milk and cottage cheese samples were collected from Oromia, Sidama, and Amhara regions. Samples were tested for Salmonella using the ISO 6579-1: 2008 method, followed by PCR confirmation. A chi-square test was conducted to assess the significance of differences in the prevalence of Salmonella in the samples between the two seasons. Results from this study showed a higher prevalence of Salmonella in all sample types during the dry season (P < 0.05). Moreover, when comparing raw milk, pasteurized milk, and cottage cheese samples, a significant difference was observed in Salmonella prevalence from raw milk samples (27.08%) collected in the Oromia region. Additionally, data showed a significantly higher prevalence of Salmonella in samples collected from raw milk producers (29.17%) during the wet season (P < 0.05). This study indicates that in order to enhance the safety of dairy products in Ethiopia, comprehensive, long-term awareness building on hygienic milk production and handling that consider seasonal influence is warranted. Supplementary Information: The online version contains supplementary material available at 10.1186/s40550-024-00108-4.

High rate of high-risk human papillomavirus among benign and breast cancer patients in Ethiopia.

INTRODUCTION: There have been numerous studies that showed the presence of human papillomavirus (HPV) in breast cancer; nonetheless, there is ongoing debate regarding their association. Given few studies in Ethiopia, we aimed to investigate the magnitude of HPV infection in Ethiopian breast cancer patients. METHODS: A total of 120 formalin-fixed paraffin-embedded (FFPE) tissue blocks were obtained, and basic demographic, clinical, and histological data were collected from medical records. DNA was extracted from archived FFPE breast tissue specimens using GeneRead DNA FFPE Kit. The AnyplexTM II HPV28 Detection Kit (Seegene, Korea) was used to detect HPV by following the manufacturer's instructions. The SPSS Version 25 was used to enter and analyze data. RESULTS: Among the 120 study participants; HPV (both high-risk and low-risk) was detected in 20.6% of breast cancer and 29.6% of non-malignant breast tumors. The most common genotype was the high-risk HPV 16 genotype. The frequency of HPV was nearly 10-fold higher in estrogen receptor-positive than ER-negative breast cancer. The percentage of HPV in the luminal (luminal A and luminal B) breast cancer subtypes was also much higher than in the non-luminal subtypes (HER-2 enriched and triple-negative breast cancer). CONCLUSION: This study did not find a significant difference in HPV expression between breast cancer and non-malignant breast tumors; however, the higher percentage of HPV in ER-positive compared to ER-negative breast cancer warrants further attention.

Genomic characterization of Listeria monocytogenes and Listeria innocua isolated from milk and dairy samples in Ethiopia.

Listeriosis caused by Listeria monocytogenes often poses a significant threat to vulnerable populations. Dairy products have been implicated in outbreaks of listeriosis worldwide. In Ethiopia, studies have identified Listeria spp. and L. monocytogenes in various dairy products, but the genetic diversity and phylogenetic relationships of these bacteria remain largely unknown in the low- and middle-income countries. Therefore, we conducted whole-genome sequencing on 15 L. monocytogenes and 55 L. innocua isolates obtained from different levels of the dairy supply chains across three regions in Ethiopia. Genomes were assembled and used for MLST genotyping and single nucleotide polymorphism (SNP) analysis to infer phylogenetic relationships. We identified a total of 3 L. monocytogenes (i.e., 2, 145, and 18) and 12 L. innocua (i.e., 1489, 1619, 603, 537, 1010, 3186, 492, 3007, 1087, 474, 1008, and 637) MLST sequence types among the studied isolates. Some of these sequence types showed region-specific occurrence, while others were broadly distributed across regions. Through high-quality SNP analysis, we found that among 13 L. monocytogenes identified as ST 2, 11 of them were highly similar with low genetic variation, differing by only 1 to 10 SNPs, suggesting potential selection in the dairy food supply chain. The L. innocua isolates also exhibited low intra-ST genetic variation with only 0-10 SNP differences, except for the ST 1619, which displayed a greater diversity.

Detection and antibiogram profile of diarrheagenic Escherichia coli isolated from two abattoir settings in northwest Ethiopia: a one health perspective.

BACKGROUND: Diarrheagenic Escherichia coli (E. coli) is a zoonotic pathogen that contaminates abattoir workers, slaughter environments, slaughter equipment, and carcasses during abattoir processing. Infection with E. coli is associated with the consumption of contaminated food and water, and it is a potential threat to the health and welfare of both humans and animals. Hence, this study aimed to detect diarrheagenic E. coli and assess its antibiogram profile in two abattoir settings, in one health lens. METHODS: A cross-sectional study in one health approach was conducted from December 2020 to June 2021. A total of 384 samples from abattoir workers' hands, carcasses, knives, cattle feces, abattoir water and effluents were collected. Bacterial culture and biochemical tests were conducted to isolate E. coli, while conventional polymerase chain reaction was performed to identify virulence genes. The antibiogram of diarrheagenic E. coli was tested against nine antimicrobials using the Kirby Bauer disk diffusion method. RESULTS: A total of 115 (29.95%) E. coli were isolated from the 384 samples, and from these isolates, about 17 (14.8%) were confirmed to be diarrheagenic E. coli (DEC). Among the DEC pathotypes, nine (52.94%), five (29.4%), and three (17.65%) were Shiga toxin-producing, enterohemorrhagic, and enterotoxigenic E. coli, respectively. While 14 (82.35%) DEC isolates harbored the stx2 gene, five (29.41%) the eae gene, five (29.41%) the hlyA gene and three (17.65%) harbored the st gene. All the DEC isolates were resistant to erythromycin and vancomycin; whereas, they were susceptible to ampicillin, nalidixic acid and norfloxacin. Furthermore, 64.7% of DEC isolates showed resistance to both ceftazidime and kanamycin and 88.24% of the isolates showed multidrug resistance. CONCLUSION: This study detected DEC isolates having different virulence genes, which showed single and multiple antimicrobial resistance. Given the existing poor hygienic and sanitary practices along the abattoir-to-table food chain, coupled with the habit of raw meat consumption, this result indicates a potential public and animal health risk from the pathogen and antimicrobial resistance.

Seroepidemiology of Toxoplasma gondii infection in women of child-bearing age in central Ethiopia.

BACKGROUND: Toxoplasma gondii infections during pregnancy can result in abortion or congenital defects. Prevalence and risk factors of toxoplasmosis in women of child-bearing age in Ethiopia are unknown. The current study was conducted with the objectives of estimating the seroprevalence and potential risk factors in acquiring T. gondii infection by women of child-bearing age in Central Ethiopia. METHODS: A cross-sectional study was conducted from March 2011 to September 2011. Sera of 425 women were analyzed by indirect enzyme linked immunosorbent assay (ELISA). A questionnaire survey was administered for all study participants to gather information on risk factors. RESULTS: The study revealed that anti- T. gondii IgG antibodies were detected in 81.4% of the samples of which 78.4% were positive for only IgG and 3.06% positive for both IgG and IgM antibodies. Seroprevalence of IgM antibodies to T. gondii (4.0%, 95% CI: 2.14, 5.86) was suggestive of recent infections. Of the 213 pregnant women 9 (4.2 %) were IgM reactive. Out of 17 potential risk factors investigated, univariate logistic regression showed significant association of T. gondii infection with study area, age, pregnancy status, raw vegetable consumption, source of water, presence of cats at home, contact with cats, HIV status and precaution during cats' feces cleaning (P ≤ 0.05). The final logistic regression model revealed that: the probability of acquiring T. gondii infection by women of Debre-Zeit was 4.46 times (95% CI of adjusted odds ratio [aOR]: 1.67, 11.89; P =0.003) higher compared to women of Ambo, pregnant women were twice (95% CI aOR: 1.13, 3.59; P = 0.018) more likely to be seropositive than non-pregnant women and women who consume raw vegetable were at increased risk of infection (aOR = 2.21, 95% CI: 1.03, 4.78; P = 0.043) than women who didn't consume. CONCLUSION: The seroprevalence of T. gondii infection in women of child-bearing age in Central Ethiopia is high. Study area, pregnancy and raw vegetable consumption are risk factors to acquire T. gondii infection. Educational program, antenatal screening of pregnant women and further epidemiological studies to uncover the economic and health impact of toxoplasmosis are suggested.

Seroepidemiological study of ovine toxoplasmosis in East and West Shewa Zones of Oromia Regional State, Central Ethiopia.

BACKGROUND: Toxoplasmosis is a globally distributed zoonosis. Consumption of raw or undercooked meat, which is among the main risk factors for acquiring human infection, is a popular tradition in Ethiopia. However, studies on toxoplasmosis in food animals used for human consumption in Ethiopia are very scarce. Thus, the objectives of the present study were to estimate the seroprevalence and the risk factors of T. gondii infection in sheep in Ambo, Ada'a-Liben and Fentale districts of Central Ethiopia. Sera from 1130 sheep were analyzed for Toxoplasma gondii specific IgG antibodies using an indirect enzyme linked immunosorbent assay (ELISA) with the P30 antigen. A questionnaire was administered to assess potential risk factors for T. gondii seropositivity. Association of seroprevalence with potential risk factors related to altitude, host and farm characteristics were analyzed by univariable and multivariable logistic regression. RESULTS: Overall flock and animal level seroprevalences were 70.48% (160/227; 95% CI: 64.51, 76.46) and 31.59% (357/1130; 95% CI: 28.88, 34.31), respectively. The multivariable logistic regression model indicated that the probability of acquiring T. gondii was higher in sheep from highland (2300 - 3200 meters above sea level) [Odds ratio (OR) = 4.11, 95% confidence interval (CI): 2.65, 6.36; P < 0.001] and midland (OR = 4.54, 95% CI: 2.76, 7.49; P < 0.001) than from lowland (<1500 meters above sea level), in females than in males (OR = 1.60, 95% CI: 1.04, 2.43, P = 0.033), in adult than in young animals (OR = 2.93, 95% CI: 1.97, 4.35, P < 0.001), in small than in large flocks (OR = 3.34, 95% CI: 1.26, 8.86, P = 0.016), and in sheep that were given tap water (OR = 4.07, 95% CI: 1.07, 15.42, P = 0.039) and river water (OR = 4.18, 95% CI: 1.54, 11.35, P = 0.005) than in those that drunk water from mixed sources (i.e., river, well, lake and pond). CONCLUSIONS: The high flock and animal level seroprevalence of toxoplasmosis in sheep is a good marker of the potential risk for human infections. Altitude, sex, age, flock size and source of water were identified as important risk factors to acquire the infection. Public education and awareness training are imperative in order to alleviate the danger posed to consumers. Further detailed studies to assess the impact of infections are warranted.

Burden of Anemia among Human Immunodeficiency Virus-Positive Adults on Highly Active Antiretroviral Therapy at Hawassa University Compressive Specialized Hospital, Hawassa, Ethiopia.

Background: Anemia is the most common hematologic abnormality associated with human immunodeficiency virus (HIV)-infected patients and affects 60% to 80% of patients in late-stage disease. It has a considerable impact on the progression of HIV to advanced stages. This study aimed at assessing the burden of anemia in adult HIV-infected patients who are on highly active antiretroviral therapy (HAART) and have follow-up at Hawassa University Comprehensive Specialized Hospital (HUCSH) Antiretroviral therapy (ART) clinic. Methods: A hospital-based retrospective study was conducted among HIV-positive adults on HAART at Hawassa University Compressive Specialized Hospital. The systematic sampling method was used to choose a total of 244 study participants. Data on demographic characteristics, related factors of anemia, latest hemoglobin, CD4, and ART regimens were collected using a structured data abstraction format. The data were cleaned and analyzed using SPSS version 21.0 after being manually checked for completeness. Multivariable logistic regression was carried out to detect elements associated with anemia. A P value of <0.05 was used as a cutoff point to announce statistical significance. Results: The records of 244 patients were examined in total. Anemia was present in 29.9% (95% CI 23.8-35.2) among adult HIV patients. Female sex (AOR: 2.576, 95% (CI: 1.295-5.127)), having tuberculosis (TB) (AOR: 4.873, 95% (CI: 1.534-15.484)), taking a zidovudine (ZDV)-containing ART regimen (AOR: 5.216, 95% (CI: 1.239-21.962)), having clinical WHO stage IV and III diseases (AOR: 3.077, 95% CI (1.244-7.612)), having body mass index (BMI) <18.5 kg/m2 (AOR: 2.391, 95% (CI: 1.138-5.023)), and taking cotrimoxazole prophylaxis (AOR: 3.860 95% (CI: 1.097-13.576)) were substantially linked to the development of anemia among adult HIV patients. Conclusion and Recommendation. This study showed that anemia is still a problem among HIV patients on HAART. The burden of anemia was found to be high among patients with advanced WHO clinical stages, having a BMI less than 18.5 kg/m2, TB/HIV coinfection, being on AZT-based ART regimens, and taking cotrimoxazole preventive therapy (CPT). Consequently, it is suggested that early preventative interventions, such as serial hemoglobin follow-up, iron supplementation, and education about dietary consumption, be undertaken targeting the aforementioned groups. In addition, the preferred first-line ART regimen as per the latest national and WHO guidelines is recommended, especially for the above groups.

Loop mediated isothermal amplification as a molecular diagnostic assay: Application and evaluation for detection of Enterohaemorrhagic Escherichia coli (O157:H7).

Purpose: This study aimed at evaluating the performance of the Loop Mediated Isothermal Amplification (LAMP) diagnostic test, which targets the putative Fimbria protein-encoding gene (Z3276) for rapid and specific detection of locally isolated enterohemorrhagic Escherichia coli (EHEC) O157:H7. Results: A total number of 40 locally available bacteria isolates and standard strains, among them 6 entrohemorrhagic (O157:H7) and 10 entropathogenic E. coli, 7 non diarrheic E. coli strains and 13 non entrohemorrhagic shiga toxic (stx) E. coli isolates as well as 4 pathogenic non E. coli species were used to optimize and evaluate the LAMP assay. The LAMP amplified DNA samples were visualized as turbid DNA both by naked eye and gel electrophoresis followed by staining. The assay had a sensitivity of 100% (6/6), a specificity of 97.05% (33/34), and an efficiency of 97.5% (39/40). The assay was also exhibited with 100% negative predicted value and 85.7% positive predicted value. The LAMP assay was also 10-fold more sensitive than the conventional PCR assay; sensitivity was determined by serial dilution. The results of LAMP and the PCR tests showed very high agreement (k = 0.97) in the detection of the bacteria studied. Conclusion: Compared with the performance of PCR and SMAC, LAMP assay was better in terms of efficiency, rapidity and cost-effectiveness, which can be used as a point-care diagnostic test in resource-limited laboratories.

Clinicopathological Features of Invasive Breast Cancer: A Five-Year Retrospective Study in Southern and South-Western Ethiopia.

Background: Breast cancer (BC) is the most common type of cancer in Ethiopia. The incidence of BC is also rising, but the exact figure is still poorly known. Therefore, this study was conducted to address the gap in epidemiological data on BC in southern and southwestern Ethiopia. Materials and Methods: This is a five-year (2015-2019) retrospective study. The demographic and clinicopathological data were collected from biopsy reports of different kinds of breast carcinomas in the pathology department of Jimma University Specialized Hospital and Hawassa University Specialized Referral Hospital. Histopathological grades and stages were conducted using Nottingham grading and TNM staging system, respectively. Collected data were entered and analyzed using SPSS Version-20 software. Results: The mean age of patients at diagnosis was 42.27 (SD = 13.57) years. The pathological stage of most BC patients was stage III, and most of them had tumor sizes greater than 5 cm. Most patients had moderately differentiated tumor grade, and mastectomy was the most common type of surgery at the time of diagnosis. Invasive ductal carcinoma was the most common histological type of BC, followed by invasive lobular carcinoma. Lymph node involvement was seen in 60.5% of cases. Lymph node involvement was associated with tumor size (χ2 = 8.55, p = 0.033) and type of surgery (χ2 = 39.69, p < 0.001). Conclusions: This study showed that BC patients in southern and southwestern Ethiopia displayed advanced pathological stages, relatively young age at diagnosis, and predominant invasive ductal carcinoma histological patterns.

High burden of ESBL and carbapenemase-producing gram-negative bacteria in bloodstream infection patients at a tertiary care hospital in Addis Ababa, Ethiopia.

BACKGROUND: Bloodstream infection due to beta-lactamase and carbapenemase-producing gram-negative bacteria poses a substantial challenge to the effectiveness of antimicrobial treatments. Therefore, this study aimed to investigate the magnitude of beta-lactamase, carbapenemase-producing gram-negative bacteria, and associated risk factors of bloodstream infections in patients at a tertiary care hospital, in Addis Ababa, Ethiopia. METHODS: An institutional-based cross-sectional study was conducted with convenience sampling techniques from September 2018 to March 2019. Blood cultures were analyzed from 1486 bloodstream infection suspected patients across all age groups. The blood sample was collected using two BacT/ALERT blood culture bottles for each patient. Gram stain, colony characteristics, and conventional biochemical tests were used to classify the gram-negative bacteria at the species level. Antimicrobial susceptibility testing was carried out to screen beta-lactam and carbapenem drug-resistant bacteria. The E-test was conducted for extended-spectrum-beta-lactamase and AmpC-beta-lactamase-producers. A modified and EDTA-modified carbapenem inactivation method was conducted for carbapenemase and metallo-beta-lactamases producers. Data collected using structured questionnaires and medical records were reviewed, encoded, and cleaned using EpiData V3.1. software. The cleaned data were exported and analyzed using SPSS version 24 software. Descriptive statistics and multivariate logistic registration models were used to describe and assess factors associated with acquiring drug-resistant bacteria infection. A p-value <0.05 was considered statistically significant. RESULT: Among 1486 samples, 231 gram-negative bacteria were identified; of these, 195(84.4%) produce drug-hydrolyzing enzymes, and 31(13.4%) produce more than one drug-hydrolyzing enzyme. We found 54.0% and 25.7% of the gram-negative bacteria to be extended-spectrum-beta-lactamase and carbapenemase-producing, respectively. The extended-spectrum-beta-lactamase plus AmpC-beta-lactamase-producing bacteria account for 6.9%. Among the different isolates Klebsiella pneumonia 83(36.7%) was the highest drug-hydrolyzing enzyme-producing bacteria. Acinetobacter spp 25(53.2%) was the most carbapenemase producer. Extended-spectrum-beta-lactamase and carbapenemase-producing bacteria were high in this study. A significant association between age groups and extended-spectrum-beta-lactamase producer bacterial infection was seen, with a high prevalence in neonates (p = <0.001). Carbapenemase showed a significant association with patients admitted to the intensive care unit (p = 0.008), general surgery (p = 0.001), and surgical intensive care unit (p = 0.007) departments. Delivery of neonates by caesarean section, and insertion of medical instruments into the body were exposing factors for carbapenem-resistant bacterial infection. Chronic illnesses were associated with an extended-spectrum-beta-lactamase-producing bacterial infection. Klebsiella pneumonia and Acinetobacter species showed the greatest rates of extensively drug-resistant (37.3%) and pan-drug-resistance (76.5%), respectively. According to the results of this study, the pan-drug-resistance prevalence was found to be alarming. CONCLUSION: Gram-negative bacteria were the main pathogens responsible for drug-resistant bloodstream infections. A high percentage of extended-spectrum-beta-lactamase and carbapenemase-producer bacteria were found in this study. Neonates were more susceptible to extended-spectrum-beta-lactamase and AmpC-beta-lactamase-producer bacteria. Patients in general surgery, caesarean section delivery, and intensive care unit were more susceptible to carbapenemase-producer bacteria. The suction machines, intravenous lines, and drainage tubes play an important role in the transmission of carbapenemase and metallo-beta-lactamase-producing bacteria. The hospital management and other stakeholders should work on infection prevention protocol implementation. Moreover, special attention should be given to all types of Klebsiella pneumoniae and pan-drug resistance Acinetobacter spp transmission dynamics, drug resistance genes, and virulence factors.