Zoonotic diseases risk perceptions and protective behaviors of consumers associated with consumption of meat and milk in and around Bishoftu, Ethiopia.

Consumption of raw or undercooked milk and meat is a major source of zoonoses. Information on the public risk perceptions and protective behaviors are essential for prevention and control of these diseases. However, such information is limited in Ethiopia. We assessed the risk perceptions and protective behaviors of the community toward zoonoses associated with consumption of raw meat and milk by employing the basic constructs of food safety health belief model. We collected data from 210 respondents (105 from each urban and peri-urban) using a face-to-face questionnaire interview. Majority of the respondents (96.2%) had knowledge about zoonoses. Despite their knowledge, 91% and 94.3% had raw meat and raw milk consumption habits, respectively. The odds of raw meat consumption was significantly higher in males (AOR = 3.90; CI = 1.28-11.86) and raw milk consumption was higher in females (AOR = 3.82; CI = 0.78-18.65). About 65% of respondents, obtained meat from backyard slaughtering. Self-owned and smallholder dairy farms are the major sources of milk for peri-urban and urban dwellers, respectively. While 46.7% of the respondents reported that community members are the primary sources of information, only 2.4% reported animal health professionals as their primary source of information. More than half of the respondents either moderately or strongly agreed that consumption of raw meat and raw milk can expose them to zoonoses. Urban dwellers had perceived more seriousness of consuming raw animal products. Peri-urban residents had significantly lower intention than urban ones to implement protective behaviors such as stopping consuming raw meat (P = 0.017) and milk (P = 0.043). We noted that lack of access to refrigerator and pasteurized milk were the perceived barriers for protection against zoonoses among the peri-urban dwellers. There was significant difference in perceived benefits of avoiding consumption of raw meat (P = 0.005) and milk (P = 0.001) between urban and peri-urban residents. Our study showed that irrespective of knowledge about zoonoses, consumptions of raw meat and raw milk remained common practices among the respondents. Public health education on the risk of consumption of raw milk and meat and the significance of protective behaviors using a one-health approach is critically needed to ensure meat and milk safety.

Investigation on Salmonella enterica, Escherichia coli, and coliforms in beef from Ethiopian abattoirs: A potential risk of meat safety.

The aims of this study were to determine the prevalence of Salmonella enterica (S. enterica) antimicrobial resistance based upon phenotypic assessment, and level of sanitation indicator organisms from 150 beef carcasses collected from three representative abattoirs in eastern, central, and southern Ethiopia. Samples were screened for S. enterica prevalence following the U.S. Department of Agriculture Microbiology Laboratory Guidebook (MLG) and confirmed by real-time PCR. The S. enterica isolates were phenotypically evaluated for susceptibility to a panel of 13 antimicrobials using disk diffusion method as described in Clinical and Laboratory Standard Institutes guidelines. In addition, the coliform, total coliform, and generic E. coli populations were quantified by plating onto E. coli/coliform petrifilms. A total of 45 isolates of S. enterica were confirmed using real-time PCR. The overall prevalence of S. enterica at the carcass level was 22.7% (95% CI, 16.0-30.0). However, based on the sampling points, S. enterica prevalence detected on hides was 13.3% (95% CI, 8.0-18.7), pre-evisceration was 12.0% (95% CI, 7.3-17.3), and post-evisceration was 4.7% (95% CI, 1.3-8.0). The prevalence of S. enterica was significantly lower on post-evisceration as compared to hides (p = .009) and pre-evisceration (p = .022). No differences were detected for the prevalence of S. enterica between abattoirs (p = .346). Finally, the prevalence of S. enterica was highest during the wet season (p = .011). The overall mean log CFU/cm2 ± SD of generic E. coli, coliform, and total coliform counts were 4.55 ± 0.99, 4.91 ± 1.13, and 4.98 ± 1.09, respectively. About 20% of S. enterica exhibited phenotypic multidrug resistant. The most frequently detected resistance was to Tetracycline (28.9%), followed by Streptomycin (22.2%) and Sulfisoxazole (20.0%). The data from this study highlight the need to implement interventions on improvement of sanitary practices in abattoirs.

One Health, "Disease X" & the challenge of "Unknown" Unknowns.

The emergence of SARS-CoV-2 and its rapid spread globally emphasizes the ever-present threat of emerging and re-emerging infectious diseases. In this review, the pathogen pyramid framework was utilized to identify the "unknown unknowns" associated with the emergence and rapid transmission of novel infectious disease agents. Given that the evolutionary origin of most of the emerging infectious disease agents can be traced to an animal source, we argue the need to integrate the "One Health" approach as a part of surveillance activities. The need for focusing on undertaking global and regional mapping activities to identify novel pathogens is discussed, given that there are an estimated 1.67 million unknown viruses, of which around 631,000 to 827,000 unknown viruses have the capacity to infect human beings. The emerging risks due to the ever-expanding interface between human, animals, both domestic and wildlife, and the environment are highlighted, these are largely driven by the need for safe habitation, growing food, developing infrastructure to support the increasing human population and desire for economic growth. The One Health approach provides a holistic way to address these cross-sectoral issues, by bridging institutional gaps, enumerating priority risk areas and pathogens, and highlighting putative risk factors for subsequent spillover events involving emerging and re-emerging infectious disease pathogens at the human-animal-environment interface.

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.

Campylobacter Colonization, Environmental Enteric Dysfunction, Stunting, and Associated Risk Factors Among Young Children in Rural Ethiopia: A Cross-Sectional Study From the Campylobacter Genomics and Environmental Enteric Dysfunction (CAGED) Project.

Livestock farming provides a possible mechanism by which smallholder farmers can meet their household need for animal source foods (ASF), which may reduce the risk of stunting. However, direct/indirect contacts with domestic animals may increase colonization by Campylobacter spp., which has been associated with Environmental Enteric Dysfunction (EED) and stunting. A cross-sectional study involving 102 randomly selected children between 12 and 16 months of age was conducted in rural eastern Ethiopia to establish prevalence rates of Campylobacter colonization, EED, and stunting, and evaluate potential risk factors. Data were collected between September and December 2018. The prevalence of EED and stunting was 50% (95% CI: 40-60%) and 41% (95% CI: 32-51%), respectively. Among enrolled children, 56% had consumed some ASF in the previous 24 h; 47% had diarrhea and 50% had fever in the past 15 days. 54, 63, 71 or 43% of households owned at least one chicken, cow/bull, goat, or sheep; 54 (53%) households kept chickens indoors overnight and only half of these confined the animals. Sanitation was poor, with high levels of unimproved latrines and open defecation. Most households had access to an improved source of drinking water. The prevalence of Campylobacter colonization was 50% (95% CI: 41-60%) by PCR. In addition to the thermotolerant species Campylobacter jejuni, Campylobacter coli and Campylobacter upsaliensis, non-thermotolerant species related to Campylobacter hyointestinalis and Campylobacter fetus were frequently detected by Meta-total RNA sequencing (MeTRS). Current breastfeeding and ASF consumption increased the odds of Campylobacter detection by PCR, while improved drinking water supply decreased the odds of EED. No risk factors were significantly associated with stunting. Further studies are necessary to better understand reservoirs and transmission pathways of Campylobacter spp. and their potential impact on child health.

Strongyloidiasis in Ethiopia: systematic review on risk factors, diagnosis, prevalence and clinical outcomes.

BACKGROUND: Strongyloidiasis is a gastrointestinal infection caused by the parasitic nematode Strongyloides stercoralis. It is estimated to infect up to 370 million people globally and is predominately found in tropical and subtropical areas of socioeconomic disadvantage. MAIN BODY: This systematic literature review identified studies published in the last ten years on the risk factors, diagnosis, prevalence and/or clinical outcomes of strongyloidiasis in Ethiopia. The prevalence of S. stercoralis ranged from 0.2 to 11.1% in adults, 0.3% to 20.7% in children, 1.5% to 17.3% in HIV positive adults and 5% in HIV positive children. The identified studies primarily used microscopy based techniques that potentially underestimated the prevalence four fold compared with serology and PCR. Strongyloidiasis in children presents a particularly significant issue in Ethiopia as children often presented with anemia, which is associated with impaired mental and cognitive development. The most significant risk factor for strongyloidiasis was HIV status and although other risk factors were identified for helminth infections, none were statistically significant for S. stercoralis specifically. Several studies detected S. stercoralis in dogs and non-biting cyclorrhaphan flies. However, future research is needed to explore the role of these reservoirs in disease transmission. CONCLUSIONS: This review demonstrated that strongyloidiasis is an overlooked and neglected disease in Ethiopia. There is a need for a systematic approach using a combination of molecular and serology based diagnostic methods to ascertain the true incidence and burden of strongyloidiasis in Ethiopia. Further research is also needed to break the cycle of transmission by identifying environmental reservoirs, risk factors and exploring the potential for zoonotic transfer.

Bovine tuberculosis in eastern Ethiopia: prevalence, risk factors and its public health importance.

BACKGROUND: Bovine tuberculosis is among the primary zoonotic disease caused by Mycobacterium bovis which has significant impact on the health of livestock and human. It has been significantly a cause for great economic loss in animal production. METHODOLOGY: A cross-sectional study was conducted from December 2014 to June 2016 on 315 cattle in selected areas of eastern Ethiopia, aiming to estimate the occurrence of bovine tuberculosis using comparative intradermal tuberculin skin test and assess cattle owners' awareness on its public health implication. Random sampling method was applied in order to select animals from farm/household and associated risk factors were recorded before purified protein derivative (PPD) injection. Forty three farm/household owners of tuberculin tested animals were interviewed using pre-tested structured questionnaires. RESULTS: The overall prevalence of bovine tuberculosis was 20.3% (n = 64) in dairy cattle at recommended cut off > 4 mm. From a total of 43 farms/households tested, 22 were positive; each farm exhibited at least one tuberculin positive reactor animal with a total herd level prevalence of 51.2%. The prevalence of bovine tuberculosis in individual animal level was significantly different (χ2 = 45.2; P-value = 0.000) in different sites with a higher prevalence (50%) in Dire Dawa. Farming system, herd size and other risk factors were significantly (p < 0.05) associated with bovine tuberculosis occurrence. Of the total interviewed farm owners, only 33% had the knowledge of or had heard about bovine tuberculosis and 23% respondents were aware of the zoonotic importance of the disease. More than 50% of the interviewees had shown their preference of raw milk consumption. Out of the total interviewed households, 3 (7%) farm workers had TB cases that had direct contact with the animals. CONCLUSION: The study showed bovine tuberculosis is highly prevalent. Associated risk factors contributed to the prevalence of the disease in cattle and its transmission. Moreover, the majority of cattle owners lack awareness about the disease and its public health significance. Awareness rising about the disease, its transmission and zoonotic implication is of great importance for reduction and control measures. Evidence of tuberculosis patient farm attendants calls also for further detail investigation.

Prevalence and Antimicrobial Susceptibility Profile of Salmonella Serovars Isolated from Slaughtered Cattle in Addis Ababa, Ethiopia.

Salmonella is one of the top causes of foodborne bacterial illnesses in humans. The primary sources of human Salmonella infection are food producing animals such as cattle, poultry, and swine. A cross-sectional study was undertaken to estimate the prevalence and to determine the serovar distribution and antimicrobial susceptibility profiles of Salmonella spp. isolated from fecal (n=567) and carcass swab (n=159) samples of slaughtered cattle at Addis Ababa Abattoir Enterprise and Kara'alo PLC, Abattoirs, in Addis Ababa, Ethiopia between January 2014 and April 2015. Salmonella isolation was conducted according to Global Foodborne Infections Network Laboratory Protocol and isolates were confirmed by genus specific PCR and serotyped by slide agglutination test. Susceptibility of the isolates to 17 antimicrobials was testedusing the Kirby-Bauer disk diffusion method according to the guidelines of the Clinical and Laboratory Standards Institute. Out of the total 726 samples examined, 27 (3.7%) were positive for Salmonella. Salmonella was detected in 4.1% (23/567) fecal and 2.5% (4/159) carcass swab samples. Twelve different serovars were identified and the most predominant serovars were S. Dublin (n=10, 35.7%) and S. Virchow (n=5, 17.9%), followed by S. Braendrerup, S. Haifa, and S. Saintpaul which were isolated from 2 samples each (7.1%). All of the Salmonella isolates investigated were resistant or intermediately resistant to four or more of the 17 drugs tested. High resistance rate was recorded to streptomycin 25 (89.3%), cephalothin 20 (71.4%), ampicillin 19 (67.9%), and amoxicillin+clavulanic acid 19 (67.9%). Resistance to five or more antimicrobials was detected in 20 (71.5%) of the isolates. Multidrug resistance to more than 7 antimicrobials was detected in 5 (17.9%) of the isolates. Isolation of such multidrug resistant strains of Salmonella from slaughtered cattle poses a major public health concern. These findings imply the need for a strict biosecurity and regulation of antimicrobial use across the country.

Brucellosis and associated risk factors in dairy cattle of eastern Ethiopia.

Knowing the status of Bovine brucellosis and associated risk factors is a crucial step in formulating evidence based control scheme. In this study, a total of 967 dairy cows from 307 dairy farms in eastern Ethiopia were serologically tested for Brucella antibodies. The screening was done first using RBPT and positive samples were subsequently subjected to CFT for confirmation. A pre-tested structured questionnaire was used to collect relevant data from 307 dairy cattle owners or attendants to assess their awareness and routine practice. The data were run using univariable logistic regression analysis using STATA version 11.0 for Windows. Accordingly, herd and individual animal seroprevalence were found to be 6.8% (95% CI = 4.28-10.28) and 1.3% (95% CI = 0.72-2.29), respectively. The prevalence of sero-reactors among local breeds was observed to be higher compared to cross breed (p < 0.05). Herd level analysis of the risk factors indicated that in farms with large herd size (>20 animals), the odds ratio (OR) = 9.13, p = 0.00, CI = 3.01-27.69 of having brucellosis was 9.13 times higher than smaller size herds (<20 animals). Intensively managed herds had shown the highest seroprevalence (20.8%) than extensive (6.7%) and semi-intensive (4.2%). Experience of dairy handlers about the disease that cause abortion in late pregnancy was significantly associated (p < 0.001) with the occurrence of brucellosis in the herds. However, about 91% of the dairy cattle owners/attendants lack awareness about disease(s) that causes abortion in late pregnancy. Similarly risk of having brucellosis in those herds experiencing abortion was 6.3 times higher (OR = 6.3, p < 0.001, CI = 2.50-15.92). This study identified some of the handling practices for aborted and retained fetal materials to be risky. Therefore, the study highlights the need of comprehensive brucellosis surveillance in animal and human and institutions of public education and on farm biosafety measures in shaping proper disease control scheme.

Reappraisal of Hydatigera taeniaeformis (Batsch, 1786) (Cestoda: Taeniidae) sensu lato with description of Hydatigera kamiyai n. sp.

The common cat tapeworm Hydatigera taeniaeformis is a complex of three morphologically cryptic entities, which can be differentiated genetically. To clarify the biogeography and the host spectrum of the cryptic lineages, 150 specimens of H. taeniaeformis in various definitive and intermediate hosts from Eurasia, Africa and Australia were identified with DNA barcoding using partial mitochondrial cytochrome c oxidase subunit 1 gene sequences and compared with previously published data. Additional phylogenetic analyses of selected isolates were performed using nuclear DNA and mitochondrial genome sequences. Based on molecular data and morphological analysis, Hydatigera kamiyai n. sp. Iwaki is proposed for a cryptic lineage, which is predominantly northern Eurasian and uses mainly arvicoline rodents (voles) and mice of the genus Apodemus as intermediate hosts. Hydatigera taeniaeformis sensu stricto (s.s.) is restricted to murine rodents (rats and mice) as intermediate hosts. It probably originates from Asia but has spread worldwide. Despite remarkable genetic divergence between H. taeniaeformis s.s. and H. kamiyai, interspecific morphological differences are evident only in dimensions of rostellar hooks. The third cryptic lineage is closely related to H. kamiyai, but its taxonomic status remains unresolved due to limited morphological, molecular, biogeographical and ecological data. This Hydatigera sp. is confined to the Mediterranean and its intermediate hosts are unknown. Further studies are needed to classify Hydatigera sp. either as a distinct species or a variant of H. kamiyai. According to previously published limited data, all three entities occur in the Americas, probably due to human-mediated introductions.

Phylogenetic characterisation of Taenia tapeworms in spotted hyenas and reconsideration of the "Out of Africa" hypothesis of Taenia in humans.

The African origin of hominins suggests that Taenia spp. in African carnivores are evolutionarily related to the human-infecting tapeworms Taenia solium, Taenia saginata and Taenia asiatica. Nevertheless, the hypothesis has not been verified through molecular phylogenetics of Taenia. This study aimed to perform phylogenetic comparisons between Taenia spp. from African hyenas and the congeneric human parasites. During 2010-2013, 233 adult specimens of Taenia spp. were collected from 11 spotted hyenas in Ethiopia. A screening based on short DNA sequences of the cytochrome c oxidase subunit 1 gene classified the samples into four mitochondrial lineages designated as I-IV. DNA profiles of nuclear genes for DNA polymerase delta (pold) and phosphoenolpyruvate carboxykinase (pepck) showed that lineages II and III can be assigned as two independent species. Common haplotypes of pold and pepck were frequently found in lineages I and IV, suggesting that they constitute a single species. Morphological observations suggested that lineage II is Taenia crocutae, but the other lineages were morphologically inconsistent with known species, suggesting the involvement of two new species. A phylogenetic tree of Taenia spp. was reconstructed by the maximum likelihood method using all protein-coding genes of their mitochondrial genomes. The tree clearly demonstrated that T. crocutae is sister to T. saginata and T. asiatica, whereas T. solium was confirmed to be sister to the brown bear tapeworm, Taenia arctos. The tree also suggested that T. solium and T. arctos are related to two species of Taenia in hyenas, corresponding to lineages I+IV and III. These results may partially support the African origin of human-infecting Taenia spp., but there remains a possibility that host switching of Taenia to hominins was not confined to Africa. Additional taxa from African carnivores are needed for further testing of the "Out of Africa" hypothesis of Taenia in humans.

Bovine cysticercosis and its food safety implications in Harari People's National Regional State, eastern Ethiopia.

Taenia saginata cysticercosis is one of the zoonotic diseases that threaten food safety and food security, particularly in developing countries. A cross-sectional study was conducted to estimate the prevalence and cyst distribution in infected cattle, and food safety implications of Taenia saginata cysticercosis in Harari People's National Regional State, eastern Ethiopia. Post-mortem inspection of carcasses and organs of slaughtered cattle in Harar Municipal Abattoir, cyst viability tests and interviews with randomly selected meat consumers were undertaken. The post-mortem inspection showed that of the 898 local zebu cattle slaughtered for human consumption and examined for the presence of cysticerci of T. saginata, 19.7% (177/898; 95% CI = 17.2-22.5) harboured at least one cyst in the muscles or organs inspected. Of the edible anatomical sites with cysticerci, shoulder muscle, liver and heart together represented 65.4%, 66.0% and 65.4% respectively of relative prevalence, total cyst count and cyst viability. These edible sites are preferred above others by local people for preparation and consumption of raw or inadequately cooked meat dishes that are locally served as kurt, kitffo and dullet. The interviews revealed that among the 300 study participants, 182 (60.7%) had been infected by taeniosis at least once during the previous year and of these 99.0% had eaten raw or undercooked beef, the majority (88.3%) obtained from butchers assumed to provide officially inspected meat that was fit for consumption. This indicated that existing meat inspection processes were inadequate to prevent carcasses infected with T. saginata cysticerci from reaching consumers. The high prevalence of viable cysts in the edible parts of beef together with the widespread consumption of raw or undercooked beef indicated the importance of T. saginata cysticercosis as a food safety problem in eastern Ethiopia. The promotion of policies to upgrade existing meat inspection procedures and public education to ensure effective prevention of T. saginata taeniosis in humans were recommended.