Hygienic status of beef butcher shop facilities and antibiotic resistance profile of Salmonella enterica in Ethiopia.

The microbiological quality of meat is influenced by the conditions of hygiene prevailing during production and handling. Thus, this study aimed to assess the prevalence of Salmonella enterica and its antimicrobial resistance, load of hygiene indicator bacteria including E. coli (ECC), coliforms (CC), total coliform (TCC), Enterobacteriaceae (EB) and aerobic plate count (APC), and meat handler's food safety knowledge and hygiene practices in butcher shops in two cities, Addis Ababa and Hawassa in Ethiopia, during 2020 and 2021. A total of 360 samples of beef carcasses (n = 120), knives (n = 60), chopping boards (n = 60), weighing balance (n = 60), and personnel's hands (n = 60) were randomly collected for microbial analysis. Besides, 120 participants were selected to participate in a food safety knowledge and hygiene practices assessment. The S. enterica isolates were identified by agglutination test followed by qPCR targeting invA gene. Phenotypic antimicrobial resistance profiles of S. enterica were determined using disk diffusion assays as described in CLSI. The ECC, CC, TCC, EB, and APC populations were quantified by plating onto petrifilm plates. A structured questionnaire was used to determine food safety knowledge and hygiene practices of participants. Overall prevalence of S. enterica was 16.7% (95% CI, 8.3-26.7) and location seems to have no effect (p = 0.806). Only 20% of the S. enterica were resistant to ampicillin and tetracycline. However, the majority (80%) of S. enterica isolates were susceptible to the panel of 11 antimicrobials tested. The overall mean ± SD (log CFU/cm2) of ECC, CC, TCC, EB, and APC were 4.31 ± 1.15; 4.61 ± 1.33; 4.77 ± 1.32; 4.59 ± 1.38 and 5.87 ± 1.52, respectively. No significant difference (p = 0.123) in E. coli contamination was observed between samples of beef carcasses and chopping boards. The EB contamination showed no significant difference (p > 0.05) among sample sources. The APC contamination levels on beef carcass were significantly higher (p > 0.05) than other sample sources. A total of 56% (95% CI: 46.7 - 65.0) of the participants had poor knowledge and 65% (95% CI: 56.7 - 73.3) had poor hygiene practices towards food safety. This study highlighted the poor hygiene status of butcher facilities with a potential risk of beef safety. Thus, appropriate food safety control strategies and inspection is needed at retail establishments.

Prevalence and Load of the Campylobacter Genus in Infants and Associated Household Contacts in Rural Eastern Ethiopia: a Longitudinal Study from the Campylobacter Genomics and Environmental Enteric Dysfunction (CAGED) Project.

In our previous cross-sectional study, multiple species of Campylobacter were detected (88%) in stool samples from children (12 to 14 months of age) in rural eastern Ethiopia. This study assessed the temporal fecal carriage of Campylobacter in infants and identified putative reservoirs associated with these infections in infants from the same region. The prevalence and load of Campylobacter were determined using genus-specific real-time PCR. Stool samples from 106 infants (n = 1,073) were collected monthly from birth until 376 days of age (DOA). Human stool samples (mothers and siblings), livestock feces (cattle, chickens, goats, and sheep), and environmental samples (soil and drinking water) from the 106 households were collected twice per household (n = 1,644). Campylobacter was most prevalent in livestock feces (goats, 99%; sheep, 98%; cattle, 99%; chickens, 93%), followed by human stool samples (siblings, 91%; mothers, 83%; infants, 64%) and environmental samples (soil, 58%; drinking water, 43%). The prevalence of Campylobacter in infant stool samples significantly increased with age, from 30% at 27 DOA to 89% at 360 DOA (1% increase/day in the odds of being colonized) (P < 0.001). The Campylobacter load increased linearly (P < 0.001) with age from 2.95 logs at 25 DOA to 4.13 logs at 360 DOA. Within a household, the Campylobacter load in infant stool samples was positively correlated with the load in mother stool samples (r2 = 0.18) and soil collected inside the house (r2 = 0.36), which were in turn both correlated with Campylobacter loads in chicken and cattle feces (0.60 < r2 < 0.63) (P < 0.01). In conclusion, a high proportion of infants are infected with Campylobacter in eastern Ethiopia, and contact with the mother and contaminated soil may be associated with early infections. IMPORTANCE A high Campylobacter prevalence during early childhood has been associated with environmental enteric dysfunction (EED) and stunting, especially in low-resource settings. Our previous study demonstrated that Campylobacter was frequently found (88%) in children from eastern Ethiopia; however, little is known about potential Campylobacter reservoirs and transmission pathways leading to infection of infants by Campylobacter during early growth. In the longitudinal study presented here, Campylobacter was frequently detected in infants within the 106 surveyed households from eastern Ethiopia, and the prevalence was age dependent. Furthermore, preliminary analyses highlighted the potential role of the mother, soil, and livestock in the transmission of Campylobacter to the infant. Further work will explore the species and genetic composition of Campylobacter in infants and putative reservoirs using PCR and whole-genome and metagenomic sequencing. The findings from these studies can lead to the development of interventions to minimize the risk of transmission of Campylobacter to infants and, potentially, EED and stunting.

Unravelling the reservoirs for colonisation of infants with Campylobacter spp. in rural Ethiopia: protocol for a longitudinal study during a global pandemic and political tensions.

INTRODUCTION: Undernutrition is an underlying cause of mortality in children under five (CU5) years of age. Animal-source foods have been shown to decrease malnutrition in CU5. Livestock are important reservoirs for Campylobacter bacteria, which are recognised as risk factors for child malnutrition. Increasing livestock production may be beneficial for improving nutrition of children but these benefits may be negated by increased exposure to Campylobacter and research is needed to evaluate the complex pathways of Campylobacter exposure and infection applicable to low-income and middle-income countries. We aim to identify reservoirs of infection with Campylobacter spp. of infants in rural Eastern Ethiopia and evaluate interactions with child health (environmental enteric dysfunction and stunting) in the context of their sociodemographic environment. METHODS AND ANALYSIS: This longitudinal study involves 115 infants who are followed from birth to 12 months of age and are selected randomly from 10 kebeles of Haramaya woreda, East Hararghe zone, Oromia region, Ethiopia. Questionnaire-based information is obtained on demographics, livelihoods, wealth, health, nutrition and women empowerment; animal ownership/management and diseases; and water, sanitation and hygiene. Faecal samples are collected from infants, mothers, siblings and livestock, drinking water and soil. These samples are analysed by a range of phenotypic and genotypic microbiological methods to characterise the genetic structure of the Campylobacter population in each of these reservoirs, which will support inference about the main sources of exposure for infants. ETHICS AND DISSEMINATION: Ethical approval was obtained from the University of Florida Internal Review Board (IRB201903141), the Haramaya University Institutional Health Research Ethics Committee (COHMS/1010/3796/20) and the Ethiopia National Research Ethics Review Committee (SM/14.1/1059/20). Written informed consent is obtained from all participating households. Research findings will be disseminated to stakeholders through conferences and peer-reviewed journals and through the Feed the Future Innovation Lab for Livestock Systems.

Isolation, antimicrobial susceptibility patterns, and risk factors assessment of non-typhoidal Salmonella from apparently healthy and diarrheic dogs.

BACKGROUND: Dogs are one of the important asymptomatic carriers of antimicrobial resistant and potentially pathogenic strains of Salmonella. They can harbor large bacterial load in the intestines and mesenteric lymph nodes which can be shed in their feces with the possibility of transmission to humans. Therefore, a cross-sectional study was conducted with the objectives of estimating the prevalence of non-typhoidal Salmonella, assessing the risk factors for dog's Salmonella carriage, and profiling the antimicrobial resistance pattern of Salmonella isolates among housed dogs in Harar town, Eastern Ethiopia. A total of 415 rectal swab samples were collected from randomly selected dogs. Samples were examined for non-typhoidal Salmonella using standard bacteriologic culture and biochemical tests. The disk diffusion method (Kirby-Bauer test) was employed to evaluate the isolates for their susceptibility against five antimicrobials. RESULTS: Non-typhoidal Salmonella were isolated from 26 (6.3%) of the rectal swab samples, with significantly higher occurrence in diarrheic (15.2%) than non-diarrheic (5.5%) dogs. The risk of Salmonella harboring was significantly higher in female dogs than in male dogs (OR = 2.5, p = 0.027). Dogs fecal shedding of Salmonella was relatively higher in households who used offal as a main feed type for their dogs (23.1%; 95% CI = 5-53.8) than those who used leftover food (10.1%; 95% CI = 5.7-16.1) and practiced mixed feeding system (17%; 95% CI = 7.6-30.8). Salmonella isolates showed higher resistance to ampicillin (41.7%), while all isolates were fully susceptible to gentamicin. Moreover, 58.3% of Salmonella isolates showed resistance to at least one of the tested antimicrobials. Majorities (72.7%) of the dog owners had no awareness on the risk of zoonotic salmonellosis from dog and all of the respondents use bare hand to clean dog kennel. CONCLUSION: Our study reveals the importance of both diarrheic and apparently healthy housed dogs in the harboring and shedding of antimicrobial resistant non-typhoidal Salmonella. The risk of non-typhoidal Salmonella spread among pet owners is not negligible, especially in households who use offal as main feed type. Therefore, an integrated approach such as: proper dog handling practices; continuous evaluation of antimicrobial resistance; and rational use of antimicrobials in the field of veterinary sector are necessary to tackle the problem.

Reproductive Diseases and Disorders of Female Camels: An Assessment and Pathological and Bacteriological Study in Eastern Ethiopia.

Camels are the most efficient domesticated animals in arid and semiarid areas of the world. In Ethiopia, they are the main livestock kept to sustain the livelihoods of pastoralists, as camels are used for milk and meat production and also for transportation. However, she-camel reproductive diseases are one of the major constraints for camel-producing communities. A cross-sectional study was conducted from November 2018 to December 2019 to identify and characterize pathological lesions and isolate possible bacteria associated with reproductive diseases and disorders in she-camels slaughtered at Dire Dawa and Babille municipal abattoirs. A total of 155 study animals were examined by recruiting all she-camels slaughtered during every abattoir visit. Overall, 562 reproductive organs, the ovaries, oviducts, uterus, and cervix, were examined through observation, palpation, and incision, and the animal- and organ-level pathological lesion prevalence were found to be 29% and 64.6%, respectively. Degenerative changes, inflammatory lesions (endometritis and salpingitis), growth disturbances (e.g., ovarian hypoplasia), and noninflammatory lesions (e.g., noninflammatory edema) were the identified pathological lesions. Occurrences of pathological changes among reproductive organs had differences where significantly the highest proportion (p = 0.00) was observed in the uteri. Of the 119 microbiological samples processed, 77.3% were positive for single or mixed bacterial genera, from which 7 different bacterial isolates and 14 other unidentified Gram-negative bacteria were detected. E. coli, Salmonella, and Staphylococcus spp. were the most frequently isolated organisms with 28.2%, 26.9%, and 12.8% frequencies, respectively. The result of the questionnaire survey showed 74% of the respondents had culled the she-camel at productive age because of poor reproductive performance associated with refused mating, abortion, and repeat breeding (poor conception). On the other hand, a majority of camel herders had poor to no information and access to modern veterinary services; nevertheless, they had good indigenous knowledge on how to manage reproductive abnormalities. Considering the importance of camels in our study area, further research on camel reproductive diseases and abnormalities with wider sample and epidemiology need to be conducted using molecular and hormonal assay techniques.

Co-occurrence of Campylobacter Species in Children From Eastern Ethiopia, and Their Association With Environmental Enteric Dysfunction, Diarrhea, and Host Microbiome.

High Campylobacter prevalence during early childhood has been associated with stunting and environmental enteric dysfunction (EED), especially in low resource settings. This study assessed the prevalence, diversity, abundance, and co-occurrence of Campylobacter spp. in stools from children in a rural area of eastern Ethiopia and their association with microbiome, diarrhea, and EED in children. Stool samples (n = 100) were collected from randomly selected children (age range: 360-498 days) in five kebeles in Haramaya District, Ethiopia. Diarrhea, compromised gut permeability, and gut inflammation were observed in 48, 45, and 57% of children, respectively. Campylobacter prevalence and species diversity were assessed using PCR and meta-total RNA sequencing (MeTRS). The prevalence of Campylobacter spp. in the children's stools was 50% (41-60%) by PCR and 88% (80-93.6%) by MeTRS (P < 0.01). Further, seven Campylobacter species (Campylobacter jejuni, Campylobacter upsaliensis, Campylobacter hyointestinalis, Campylobacter coli, Campylobacter sp. RM6137, uncultured Campylobacter sp., and Campylobacter sp. RM12175) were detected by MeTRS in at least 40% of children stools in high abundance (>1.76-log read per million per positive stool sample). Four clusters of Campylobacter species (5-12 species per cluster) co-occurred in the stool samples, suggesting that Campylobacter colonization of children may have occurred through multiple reservoirs or from a reservoir in which several Campylobacter species may co-inhabit. No associations between Campylobacter spp., EED, and diarrhea were detected in this cross-sectional study; however, characteristic microbiome profiles were identified based on the prevalence of Campylobacter spp., EED severity, and diarrhea. Forty-seven bacterial species were correlated with Campylobacter, and 13 of them also correlated with gut permeability, gut inflammation and/or EED severity. Forty-nine species not correlated with Campylobacter were correlated with gut permeability, gut inflammation, EED severity and/or diarrhea. This study demonstrated that (1) in addition to C. jejuni and C. coli, multiple non-thermophilic Campylobacter spp. (i.e., Campylobacter hyointestinalis, Campylobacter fetus, and Campylobacter concisus) were frequently detected in the children's stools and (2) the Campylobacter, gut permeability, gut inflammation, EED severity, and diarrhea were associated with characteristic microbiome composition. Additional spatial and longitudinal studies are needed to identify environmental reservoirs and sources of infection of children with disparate Campylobacter species and to better define their associations with EED in low-income countries.

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.

In Vitro Antibacterial Activity of Selected Medicinal Plants in the Traditional Treatment of Skin and Wound Infections in Eastern Ethiopia.

BACKGROUND: External infections involving the skin and wound are the most frequent complications affecting humans and animals. Medicinal plants play great roles in the treatment of skin and wound infections. This study was aimed to evaluate the in vitro antibacterial activity of crude methanolic extracts of nine medicinal plants. METHODS: Agar well diffusion and broth dilution methods were used to determine the antibacterial activity of nine Ethiopian plants against four bacterial species including Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. RESULTS: Among the tested plants, seven (Cissus quadrangularis, Commelina benghalensis, Euphorbia heterophylla, Euphorbia prostrate, Momordica schimperiana, Trianthema spp., and Solanum incanum) were found to exhibit considerable antibacterial activity against at least one of the test bacteria. The extracts of C. quadrangularis, E. heterophylla, and E. prostrata had a wide spectrum of antibacterial activities against test bacterial strains while the extracts of Grewia villosa and Schinus molle did not show any inhibitory activity. Clinical isolate and laboratory strain of S. aureus showed the highest susceptibility to highest concentration (780 mg/mL) of E. prostrata with a zone of inhibition of 21.0mm and 22.3mm, respectively. CONCLUSION: This study indicates clear evidence supporting the traditional use of seven plants in treating skin and wound infections related to bacteria.

Epidemiological and therapeutic studies of camel mange in Fafan zone, Eastern Ethiopia.

BACKGROUND: Camel mange is an economically important parasitic disease affecting productivity in camel rearing areas of the world if appropriate treatment is not instituted. METHODS: A cross-sectional and a controlled field trial were carried out to study the epidemiology of camel mange in Fafan zone, Eastern Ethiopia, and evaluate the efficacy of ivermectin and diazinon in the control of mange infestation in camels on the basis of clinical and parasitological evidence, respectively. Three groups of naturally infested camels and one group of healthy camels each composed of 6 individuals were enrolled: the two infested groups received either ivermectin or diazinon, and the other groups remained untreated. RESULTS: The overall prevalence rate of mange in camels in the study area was 31.5% and the only identified species was Sarcoptes scabiei. The prevalence rate was found to significantly vary (p < 0.05) in relation to body condition and herd size of camels. Both drugs showed significant variation (p < 0.05) on improving clinical and body condition scores. Clearance of mange lesions occurred with both drugs; however, re-infestation was observed in diazinon treated group. Ivermectin significantly improved (p < 0.05) both body condition and clinical scores whereas diazinon markedly improved only the later. CONCLUSION: In conclusion, camels in the study area harbored considerable level of S. scabiei which warrants institution of an integrated control approach by administration of ivermectin while also sanitating the animal environment.