Assessment of health problems of sheep and goats based on ante-mortem and post-mortem inspection at Addis Ababa Abattoir, Ethiopia.

Introduction: Ethiopia has a rapidly growing small ruminant sector, which faces low productivity due to husbandry practices and poor health condition of the animals. A study was conducted in Ethiopia's largest municipal abattoir with the objective to assess the health problems of sheep and goats presented for slaughter using standard ante-mortem and post-mortem methodology. Methods: A cross-sectional study using systematic random sampling was conducted on 384 sheep and 384 goats from January to July 2014. Results: Soiled skin (69.1%), poor body condition (24.3%), and nostril discharge (19.5%) were common among both species at ante-mortem examination. Gross lesions were frequent in livers (39.7%) and lungs (37.2%), while pneumonia (18.1%) and adhesions (13.8%) were frequent in the lungs of sheep and goats, indicating stress-related illness. Parasitic lesions, especially fasciolosis (19.3%) and hydatid cysts (8.1%) were significantly more common in sheep livers (p 0.05). The direct financial loss from lesions in both species was 1,077,015 ETB or 53,851 USD per year, most of which was estimated to occur from carcass bruising. Discussion: The findings indicate that reducing parasite burden and preventing carcass bruising through improved handling could significantly increase the profitability of the small ruminant meat sector in Ethiopia.

Assessment of Small Ruminant Welfare in Ethiopia - An Abattoir-Based Study.

This paper presents the results of a cross-sectional study assessing the welfare of sheep and goats in a large abattoir in central Ethiopia, using qualitative and quantitative approaches. A total of 384 nonhuman animals (192 sheep and 192 goats) underwent clinical examination and behavioral observation in 2014. The study also included behavioral observation of 57 animal handlers, and a qualitative assessment of animal welfare conditions at the abattoir. Clinical examination revealed dirty wool/hair (54.9%), poor body condition (15.8%), and respiratory disorders (14.0%). The most commonly observed behaviors were refusing to move (17.1%), panting (15.8%) and reversing (10.4%). Significantly higher proportion of sheep exhibited resistance behaviors such as refusing to move (21.4% of sheep and 13.0% of goats; p < 0.030) and resistance to being pulled (12.5% of sheep and 6.3% of goats; p < 0.0036). Handlers frequently beat (87.7%), pushed (57.9%) and pulled (49.1%) the animals. Poor handling of animals was very common and could result in animal distress and falls. In collaboration with the abattoir workers, the authors developed simple good practice to improve welfare and reduce losses from poor welfare.

Growth and Aflatoxin B1, B2, G1, and G2 Production by Aspergillus flavus and Aspergillus parasiticus on Ground Flax Seeds (Linum usitatissimum).

ABSTRACT: Flax seed has become an increasingly popular food ingredient because of its nutrient richness as well as potential health benefits. Flax seeds are often ground before consumption, and flax seed cakes are used as animal feed. Aflatoxin production may occur subsequently when the ground seeds are stored in an environment that supports fungal growth. The objectives of this study were to determine the growth of two toxigenic fungi, Aspergillus flavus and A. parasiticus, and to quantify the concentrations of four major aflatoxins (AFB1, AFG1, AGB2, and AFG2) produced by the two fungi on ground flax seeds with water activities (aws) of 0.82, 0.86, 0.90, 0.94, and 0.98, incubated for 30 days at 20, 27, and 35°C. Results of the study showed that A. flavus was able to grow on ground seeds with aw from 0.86 to 0.98 at all three temperatures, and the most rapid growth occurred at aws 0.90 and 0.94 at 27°C. In comparison, A. parasiticus grew on seeds with aw from 0.86 to 0.98 at 27 and 35°C as well as on seeds with aw from 0.86 to 0.90 at 20°C, and the most favorable growth condition was aw from 0.90 to 0.94 at 35°C. A. flavus produced aflatoxins on seeds with aw from 0.90 to 0.94 at 27°C as well as on seeds with aw from 0.86 to 0.98 at 35°C, and the maximum total aflatoxin (298 µg/kg), AFB1 (247 µg/kg), and AFG1 (51 µg/kg) were found on seeds with aw 0.90 at 35°C. In comparison, A. parasiticus produced aflatoxins under a wider range of conditions, which included aw 0.86 at 27 and 35°C, aw 0.90 at 20 and 27°C, aw 0.94 at 27°C, and aw 0.98 at 35°C. The maximum total aflatoxin (364 µg/kg) and maximum AFB1 (324 µg/kg) along with 34 µg/kg AFG1 and 6 µg/kg AFB2 were produced by A. parasiticus on seeds with aw 0.98 incubated at 35°C for 30 days. Linear regression models also indicated that high incubation temperature (35°C) was optimal for overall fungal growth and for formation of high levels of aflatoxin by both fungi. Future studies should also address aflatoxin contamination in flax seed oil.

Aflatoxin B1 (AFB1) production by Aspergillus flavus and Aspergillus parasiticus on ground Nyjer seeds: The effect of water activity and temperature.

Nyjer oil seed cake supports high levels of aflatoxin B1 (AFB1) production. AFB1 is a secondary metabolite of Aspergillus flavus and A. parasiticus, classified as a Class 1A carcinogen. The aim of this study was to determine the effects of temperature (20, 27, and 35 °C) and water activity (0.82, 0.86, 0.90, 0.94, and 0.98 aw) on fungal growth and AFB1 production of A. flavus and A. parasiticus on ground Nyjer seeds over a 30-day incubation period. Linear regression models indicated that both fungal growth and AFB1 production were significantly influenced by water activity of Nyjer seeds and incubation temperature. The two fungi did not grow on Nyjer seeds at 0.82 aw at the three incubation temperatures. The most favorable growth conditions for both fungi were 0.90-0.98 aw at 27 °C or 0.90-0.94 aw at 35 °C. The optimum temperature for AFB1 production was 27 °C for both A. flavus and A. parasiticus (with regression coefficients of 6.01 and 9.11, respectively). Both fungi were likely to produce high levels of AFB1 at 0.90 aw (with regression coefficients of 3.56 for A. flavus and 7.17 for A. parasiticus). Aspergillus flavus only produced AFB1 on seeds with 0.90-0.98 aw at 27 °C (in the range of 203-282 µg/kg) and on seeds with 0.90 aw at 35 °C (212 µg/kg). No detectable AFB1 was produced by this fungus in any other culture conditions that were studied. Aspergillus parasiticus, in contrast, was able to produce AFB1 under all of the growth conditions. At 20 °C, this fungus produced the highest level of AFB1 (212 µg/kg) at high water activity (0.98 aw). At 27 °C, A. parasiticus produced high levels of AFB1 (in the range of 209-265 µg/kg) at a wide range of water activities (0.86-0.98 aw). In the entire study, the highest AFB1 concertation for A. parasiticus was detected on seeds incubated at high temperature (35 °C) and low water activity (0.86 aw). The findings of this study could help optimize the storage conditions of Nyjer oil seeds to reduce aflatoxin contamination.

Milk handling practices and consumption behavior among Borana pastoralists in southern Ethiopia.

BACKGROUND: Safety and wholesomeness of milk intended for human consumption are influenced by various interlinked factors. However, information on what these factors are, especially in the pastoral traditional communities of Ethiopia, is largely lacking. The objective of this study is to assess the hygienic milk production, processing and consumption practices, and behaviors of Borana pastoralists. METHODS: The study used qualitative participatory research methods. Individual semi-structured interviews, focus group discussions, informal discussions, and observations were carried out on (1) milk handling practices, (2) perceptions of quality and safety of milk, including perceived criteria for good milk, (3) awareness of milk-borne diseases, and (4) perception towards milk boiling practices. The interviews and discussions were audio recorded, transcribed, and analyzed by identifying themes. RESULTS: Some risky behaviors related to milk handling and consumption were identified. These include unhygienic conditions in handling milk and milk products, consumption behaviors such as consuming raw milk purchased from markets, and children directly consuming milk from the udder of animals (e.g., goats). There was a very strong reluctance to boil milk before consumption mainly because of the misconception that nutrients in the milk are destroyed when milk is boiled and "boiled milk is dead". On the other hand, potential risk mitigation practices were identified such as smoking of milk containers (which may help reduce microbial growth), processing milk through fermentation, consuming milk in boiled tea, and a recent trend towards boiling milk for babies. However, the latter was not motivated by concern over microbial hazards but the belief that raw milk could form curds in children's stomach which might then suffocate them. CONCLUSION: The findings highlight the need to promote hygienic handling practices of milk and closely engage with local communities to improve their understanding of milk safety to facilitate change in practices. Educating pastoralists on good milk production practices should be given priority. One of the ways to do this could be by strengthening the integration of milk hygiene in research and development programs as an entry point for behavioral change towards the safe handling and consumption of milk and milk products.

Effect of water activity, temperature, and incubation period on fungal growth and ochratoxin A production on Nyjer seeds.

Aspergillus fresenii and Aspergillus sulphureus produce ochratoxin A (OTA), which is a secondary metabolite of Aspergillus and Penicillium species, with nephrotoxic effects and potential carcinogenic activity. The aim of this study was to determine the effects of temperature (20, 30, and 37 °C), water activity (0.82, 0.86, 0.90, 0.94, and 0.98 aw), incubation period (5, 10, 15, and 30 days) on fungal growth, and OTA production by A. fresenii and A. sulphureus on Nyjer oil seeds. There was no fungal growth at 0.82 aw. The two fungal species were able to produce OTA from the fifth day of incubation from 0.86 to 0.98 aw and temperature 20 to 37 °C. Aspergillus fresenii produced the highest concentration of OTA (643 µg/kg) at 0.90 aw and 37 °C within 15 days, while A. sulphureus produced the highest level of OTA (724 µg/kg) at 0.98 aw and 20 °C within 10 days. The optimum water activity and temperature for the growth of both fungi were similar at 0.94 aw and at 30 °C. There was statistically significant difference between the levels of OTA production among the two fungi. Overall, A. sulphureus produced significantly higher levels of OTA (p < 0.05). Higher temperature (37 °C) and 0.90-0.94 aw were optimal for OTA production by A. fresenii. Our results show that Nyjer seeds can support the growth of A. fresenii and A. sulphureus and OTA production, and the two species had similar temperature and water activity requirements for growth but different requirements for OTA production on Nyjer seeds.

Important knowledge gaps among pastoralists on causes and treatment of udder health problems in livestock in southern Ethiopia: results of qualitative investigation.

BACKGROUND: Ethiopia has high prevalences of udder health problems including clinical and subclinical mastitis across production systems in different livestock species. Previous studies on udder health problems have largely focused on identification of mastitis causing microbial pathogens and associated risk factors. However, relatively little is known about the knowledge and beliefs of livestock keepers regarding udder health problems. An understanding of the beliefs on the other hand would facilitate effective communication between livestock keepers and animal health professionals. Therefore, this study aimed at exploring the knowledge and belief surrounding the causes, clinical signs and treatments for udder health problems in (agro-) pastoral communities in southern Ethiopia using qualitative investigation. RESULTS: The result showed that udder health problem, locally known as 'dhukkuba muchaa', which translates to 'disease of teats', was classified into three main types: (1) tick infestation (dirandisa), (2) swelling of udder often with pus discharge (nyaqarsa) and (3) acute mastitis caused by evil eye (buda) with 'bloody milk'. Tick infestation was perceived to directly cause mechanical damage to udder tissue or to resulting in swelling leading to nyaqarsa. Our analysis also revealed the strong misperception that acute and severe swelling of udder was caused by evil eye. According to the pastoralists, cows with large udders in the late pregnancy are prone to evil eye infliction upon giving birth. The pastoralists often treat udder health problems by combining both modern and traditional methods. Removal of ticks by hand and acarcide application were the preferred methods for limiting tick infestation while swelling and evil eye cases were treated with antibiotics (e.g. oxytetracycline). The study also revealed that specific herbs, only known by the herbalists, were used for traditional treatment of udder health. Although this information could not be divulged at the time, it should form the subject of further investigation. Traditional treatment for evil eye was often administered through nostrils, raising questions about its effectiveness. CONCLUSION: The narration given by the pastoralists in associating tick infestation with udder health problems was compatible with existing scientific evidences. In this respect, such local knowledge can be better utilized for the educational messages targeting control and management of tick infestation in livestock. However, the misperception of causes for acute udder swelling as evil eye can be problematic as far as the application of appropriate treatment and management of the problem is concerned. The misperception can significantly impact the welfare of animals and highlights the need for capacity building of the pastoralists on the causes and treatment of udder health problems.

Major vectors and vector-borne diseases in small ruminants in Ethiopia: A systematic review.

Vector-borne diseases are among major health constraints of small ruminant in Ethiopia. While various studies on single vector-borne diseases or presence of vectors have been conducted, no summarized evidence is available on the occurrence of these diseases and the related vectors. This systematic literature review provides a comprehensive summary on major vectors and vector-borne diseases in small ruminants in Ethiopia. Search for published and unpublished literature was conducted between 8th of January and 25th of June 2015. The search was both manual and electronic. The databases used in electronic search were PubMed, Web of Science, CAB Direct and AJOL. For most of the vector-borne diseases, the summary was limited to narrative synthesis due to lack of sufficient data. Meta-analysis was computed for trypanosomosis and dermatophilosis while meta-regression and sensitivity analysis was done only for trypanososmosis due to lack of sufficient reports on dermatophilosis. Owing emphasis to their vector role, ticks and flies were summarized narratively at genera/species level. In line with inclusion criteria, out of 106 initially identified research reports 43 peer-reviewed articles passed the quality assessment. Data on 7 vector-borne diseases were extracted at species and region level from each source. Accordingly, the pooled prevalence estimate of trypanosomosis was 3.7% with 95% confidence interval (CI) 2.8, 4.9), while that of dermatophilosis was 3.1% (95% CI: 1.6, 6.0). The in-between study variance noted for trypanosomosis was statistically significant (p<0.05). Among the three covariates considered for meta-regression, only one (species) fitted the final model significantly (p<0.05) and explained 65.44% of the between studies variance (R2). The prevalence in sheep (5.5%) increased nearly by 34% compared to goats (2.9%). The parasitic presence in blood was documented for babesiosis (3.7% in goats); and anaplasmosis (3.9% in sheep). Serological evidence was retrieved for bluetongue ranging from 34.1% to 46.67% in sheep, and coxiellosis was 10.4% in goats. There was also molecular evidence on the presence of theileriosis in sheep (93%, n=160) and goats (1.9%, n=265). Regarding vectors of veterinary importance, 14 species of ticks in five genera, four species of Glossina and 4 genera of biting flies were reported. Despite the evidence on presence of various vectors including ticks, flies, mosquitoes and midges, studies on vector-borne diseases in Ethiopia are surprisingly rare, especially considering risks related to climate change, which is likely to affect distribution of vectors. Thus better evidence on the current situation is urgently needed in order to prevent spread and to model future distribution scenarios.

Systematic review and meta-analysis of metacestodes prevalence in small ruminants in Ethiopia.

Metacestodes, the larval stages of canid cestode parasites, are among the causes of morbidity, mortality and financial losses in small ruminants in Ethiopia as a result of organ and carcass condemnation at slaughter. Several studies have been conducted over the years; however, these studies often had limited scope and coverage. This systematic review and meta-analysis was conducted to collate the information so far available in order to provide a pooled prevalence estimate at national level and identify potential predictors. Published and grey literature written in English and Amharic in the period from, 1st of January 1990 to June 25, 2015 were searched from electronic databases and repositories of academic and research institutions. Relevant animal level data on 67,743 small ruminants was extracted from 23 published articles and one master's thesis resulting altogether in 86 animal level reports that conformed to predefined criteria. The dataset was analyzed using a meta-analytical approach. The pooled prevalence estimate computed for metacestodes infection was 11.8% with a 95% confidence interval (CI) of 9.1, 15.4. The highest prevalence estimate 31.2% (95% CI: 23.1, 40.9) was found for Cysticercus tenuicollis (Taenia hydatigena) followed by cystic echinococcosis (Echinococcus granulosus) 8.8% (95% CI: 5.8, 13.1), Cysticercus ovis (Taenia ovis) 4.9% (95% CI: 2.9, 9.4) and Coenurus cerebralis (Taenia multiceps) 4.6% (95% CI: 1.6, 12.2). Among the predictors considered for heterogeneity analysis only sample size and metacestode type fitted the final multivariable meta-regression model and explained 26.3% of the explainable heterogeneity between studies (p<0.05). The prevalence was noted to decrease with increasing sample size. No significant difference in prevalence was observed between sheep and goats (p>0.05). In conclusion, this review showed a widespread occurrence of metacestodes in small ruminants in Ethiopia. Thus, a holistic approach to break the life cycle of these parasitic stages is suggested, including regulatory interventions that encourage dog owners to keep their dogs confined and prevent backyard slaughter and proper management of abattoir waste disposal.

Gastrointestinal nematode infection in small ruminants in Ethiopia: A systematic review and meta-analysis.

Gastrointestinal (GI) nematode infections are a major health challenge affecting productive and reproductive performance of sheep and goats in Ethiopia. However, there is no comprehensive summary on the occurrence and distribution of the infection at national level. This systematic review provides pooled prevalence estimates and assesses potential predictors of the nematode infections in small ruminants, i.e. helpful in planning interventions or control strategies. The review used 50 animal level datasets retrieved from 24 manuscripts. The studies used data collected from 9407 sheep and 3478 goats. A meta-analytical approach was employed to analyze Effect size (ES). The reported GI nematodes represented eleven genera affecting sheep and goats including: Haemonchus, Trichostrongylus, Teladorsagia/Ostertagia, Strongyloides, Bunostomum, Nematodirus, Chabertia, Trichuris, Cooperia, Skrjabinema and Oesophagostomum. The GI nematodes pooled prevalence estimate in the random effect model was 75.8% (95% CI: 69.6, 80.8). The subgroup analysis revealed significant (p<0.05) differences in the prevalence between different regions and type of diagnostic methods used. 'Postmortem technique' and 'eastern part of the country' were associated with higher GI nematode prevalence and accounted for 68.1% of the between studies heterogeneity. In light of the high parasitic prevalence in all agro-ecologies, the need for strategic intervention is recommended. Meanwhile, data need to be generated for some of the regions where dependable survey reports are lacking.

A meta-analysis of contagious caprine pleuropneumonia (CCPP) in Ethiopia.

This systematic literature review was initiated due to lack of comprehensive information on the status and distribution of contagious caprine pleuropneumonia (CCPP) in Ethiopia. The objectives of the review were thus to provide a pooled prevalence estimate of CCPP in the country and asses the level of in between study variance among the available reports. Manual and electronic search was conducted between 8th of January and 25th of June 2015. A total of twelve published articles and one MSc thesis was retrieved from 19 initially identified studies. Twenty five animal level datasets were extracted at regional level considering some hypothesized predictors. The retrieved data were summarized in a meta-analytical approach. Accordingly, the pooled prevalence estimate of CCPP was 25.7% (95% CI:20.9,31.0). The inverse variance square (I(2)) that explains the variation in effect size attributed to reports true heterogeneity was 95.7%.The sub-group analysis was also computed for assumed predictors including, age, sex, type of study population, production systems and regional states. Among these predictors, study population type revealed statistically significant difference (P<0.05). Accordingly, the prevalence estimate for samples collected at abattoir was 39.2%, while that of samples collected at field level was 22.4%. In the final model, type of study population fitted the multivariable meta-regression model accounting for 22.87% of the explainable proportion of heterogeneity among the presumed predictors. Evidence on isolation and confirmation of Mycoplasma capricolum subspp. capripneumonie in the country was obtained from five regional states. In conclusion, it is recommended to further investigate facilities related with transportation and collection premises along with potential role of sheep in the epidemiology of CCPP. Finally, the review emphasizes the need for monitoring the ongoing CCPP control intervention and introduces amendments based on the findings. Besides more surveys are needed in some of the regions where no or few valid data was available.

Isolation of Multidrug-Resistant Escherichia coli O157 from Goats in the Somali Region of Ethiopia: A Cross-Sectional, Abattoir-Based Study.

Toxigenic Escherichia coli (E. coli) are an important cause of gastroenteritis in developing countries. In Ethiopia, gastroenteritis due to food-borne disease is a leading cause of death. Yet, there is no surveillance for E. coli O157 and little is known about the carriage of this pathogen in Ethiopia's livestock. This study aimed to assess the prevalence and levels of antimicrobial resistance of E. coli O157 in goat meat, feces, and environmental samples collected at a large abattoir in the Somali region of Ethiopia. The samples were enriched in modified tryptone broth containing novobiocin, and plated onto sorbitol MacConkey agar. Isolates were confirmed using indole test and latex agglutination. Antimicrobial susceptibility testing was conducted using the disk diffusion method. A total of 235 samples, including 93 goat carcass swabs, 93 cecal contents, 14 water, 20 hand, and 15 knife swabs were collected. Overall, six (2.5%) samples were contaminated with E. coli O157 of which two (2.1%) were isolated from cecal contents, three (3.2%) from carcass swabs, and one (7.1%) from water. All isolates were resistant to at least two of the 18 antimicrobials tested. Two isolates (33.3%) were resistant to more than five antimicrobials. Abattoir facilities and slaughter techniques were conducive to carcass contamination. This study highlights how poor hygiene and slaughter practice can result in contaminated meat, which is especially risky in Ethiopia because of the common practice of eating raw meat. We detect multi-resistance to drugs not used in goats, suggesting that drugs used to treat human infections may be the originators of antimicrobial resistance in livestock in this ecosystem. The isolation of multidrug-resistant E. coli O157 from goats from a remote pastoralist system highlights the need for global action on regulating and monitoring antimicrobial use in both human and animal populations.

Exploratory spatial analysis of Lyme disease in Texas -what can we learn from the reported cases?

BACKGROUND: Lyme disease (LD) is a tick-borne zoonotic illness caused by the bacterium Borrelia burgdorferi. Texas is considered a non-endemic state for LD and the spatial distribution of the state's reported LD cases is unknown. METHODS: We analyzed human LD cases reported to the Texas Department of State Health Services (TX-DSHS) between 2000 and 2011 using exploratory spatial analysis with the objective to investigate the spatial patterns of LD in Texas. Case data were aggregated at the county level, and census data were used as the population at risk. Empirical Bayesian smoothing was performed to stabilize the variance. Global Moran's I was calculated to assess the presence and type of spatial autocorrelation. Local Indicator of Spatial Association (LISA) was used to determine the location of spatial clusters and outliers. RESULTS AND DISCUSSION: There was significant positive spatial autocorrelation of LD incidence in Texas with Moran's I of 0.41 (p = 0.001). LISA revealed significant variation in the spatial distribution of human LD in Texas. First, we identified a high-risk cluster in Central Texas, in a region that is thought to be beyond the geographical range of the main vector, Ixodes scapularis. Second, the eastern part of Texas, which is thought to provide the most suitable habitat for I. scapularis, did not appear to be a high-risk area. Third, LD cases were reported from several counties in western Texas, a region considered unsuitable for the survival of Ixodes ticks. CONCLUSIONS: These results emphasize the need for follow-up investigations to determine whether the identified spatial pattern is due to: clustering of misdiagnosed cases, clustering of patients with an out-of state travel history, or presence of a clustered unknown enzootic cycle of B. burgdorferi in Texas. This would enable an improved surveillance and reporting of LD in Texas.

Multifactorial effects of ambient temperature, precipitation, farm management, and environmental factors determine the level of generic Escherichia coli contamination on preharvested spinach.

A repeated cross-sectional study was conducted to identify farm management, environment, weather, and landscape factors that predict the count of generic Escherichia coli on spinach at the preharvest level. E. coli was enumerated for 955 spinach samples collected on 12 farms in Texas and Colorado between 2010 and 2012. Farm management and environmental characteristics were surveyed using a questionnaire. Weather and landscape data were obtained from National Resources Information databases. A two-part mixed-effect negative binomial hurdle model, consisting of a logistic and zero-truncated negative binomial part with farm and date as random effects, was used to identify factors affecting E. coli counts on spinach. Results indicated that the odds of a contamination event (non-zero versus zero counts) vary by state (odds ratio [OR] = 108.1). Odds of contamination decreased with implementation of hygiene practices (OR = 0.06) and increased with an increasing average precipitation amount (mm) in the past 29 days (OR = 3.5) and the application of manure (OR = 52.2). On contaminated spinach, E. coli counts increased with the average precipitation amount over the past 29 days. The relationship between E. coli count and the average maximum daily temperature over the 9 days prior to sampling followed a quadratic function with the highest bacterial count at around 24°C. These findings indicate that the odds of a contamination event in spinach are determined by farm management, environment, and weather factors. However, once the contamination event has occurred, the count of E. coli on spinach is determined by weather only.

Farm management, environment, and weather factors jointly affect the probability of spinach contamination by generic Escherichia coli at the preharvest stage.

The National Resources Information (NRI) databases provide underutilized information on the local farm conditions that may predict microbial contamination of leafy greens at preharvest. Our objective was to identify NRI weather and landscape factors affecting spinach contamination with generic Escherichia coli individually and jointly with farm management and environmental factors. For each of the 955 georeferenced spinach samples (including 63 positive samples) collected between 2010 and 2012 on 12 farms in Colorado and Texas, we extracted variables describing the local weather (ambient temperature, precipitation, and wind speed) and landscape (soil characteristics and proximity to roads and water bodies) from NRI databases. Variables describing farm management and environment were obtained from a survey of the enrolled farms. The variables were evaluated using a mixed-effect logistic regression model with random effects for farm and date. The model identified precipitation as a single NRI predictor of spinach contamination with generic E. coli, indicating that the contamination probability increases with an increasing mean amount of rain (mm) in the past 29 days (odds ratio [OR] = 3.5). The model also identified the farm's hygiene practices as a protective factor (OR = 0.06) and manure application (OR = 52.2) and state (OR = 108.1) as risk factors. In cross-validation, the model showed a solid predictive performance, with an area under the receiver operating characteristic (ROC) curve of 81%. Overall, the findings highlighted the utility of NRI precipitation data in predicting contamination and demonstrated that farm management, environment, and weather factors should be considered jointly in development of good agricultural practices and measures to reduce produce contamination.

Generic Escherichia coli contamination of spinach at the preharvest stage: effects of farm management and environmental factors.

The objective of this study was to determine the effects of farm management and environmental factors on preharvest spinach contamination with generic Escherichia coli as an indicator of fecal contamination. A repeated cross-sectional study was conducted by visiting spinach farms up to four times per growing season over a period of 2 years (2010 to 2011). Spinach samples (n = 955) were collected from 12 spinach farms in Colorado and Texas as representative states of the Western and Southwestern United States, respectively. During each farm visit, farmers were surveyed about farm-related management and environmental factors using a questionnaire. Associations between the prevalence of generic E. coli in spinach and farm-related factors were assessed by using a multivariable logistic regression model including random effects for farm and farm visit. Overall, 6.6% of spinach samples were positive for generic E. coli. Significant risk factors for spinach contamination with generic E. coli were the proximity (within 10 miles) of a poultry farm, the use of pond water for irrigation, a >66-day period since the planting of spinach, farming on fields previously used for grazing, the production of hay before spinach planting, and the farm location in the Southwestern United States. Contamination with generic E. coli was significantly reduced with an irrigation lapse time of >5 days as well as by several factors related to field workers, including the use of portable toilets, training to use portable toilets, and the use of hand-washing stations. To our knowledge, this is the first report of an association between field workers' personal hygiene and produce contamination with generic E. coli at the preharvest level. Collectively, our findings support that practice of good personal hygiene and other good farm management practices may reduce produce contamination with generic E. coli at the preharvest level.

Risk factors for microbial contamination in fruits and vegetables at the preharvest level: a systematic review.

The objective of this study was to perform a systematic review of risk factors for contamination of fruits and vegetables with Listeria monocytogenes, Salmonella, and Escherichia coli O157:H7 at the preharvest level. Relevant studies were identified by searching six electronic databases: MEDLINE, EMBASE, CAB Abstracts, AGRIS, AGRICOLA, and FSTA, using the following thesaurus terms: L. monocytogenes, Salmonella, E. coli O157 AND fruit, vegetable. All search terms were exploded to find all related subheadings. To be eligible, studies had to be prospective controlled trials or observational studies at the preharvest level and had to show clear and sufficient information on the process in which the produce was contaminated. Of the 3,463 citations identified, 68 studies fulfilled the eligibility criteria. Most of these studies were on leafy greens and tomatoes. Six studies assessed produce contamination with respect to animal host-related risk factors, and 20 studies assessed contamination with respect to pathogen characteristics. Sixty-two studies assessed the association between produce contamination and factors related to produce, water, and soil, as well as local ecological conditions of the production location. While evaluations of many risk factors for preharvest-level produce contamination have been reported, the quality assessment of the reviewed studies confirmed the existence of solid evidence for only some of them, including growing produce on clay-type soil, the application of contaminated or non-pH-stabilized manure, and the use of spray irrigation with contaminated water, with a particular risk of contamination on the lower leaf surface. In conclusion, synthesis of the reviewed studies suggests that reducing microbial contamination of irrigation water and soil are the most effective targets for the prevention and control of produce contamination. Furthermore, this review provides an inventory of the evaluated risk factors, including those requiring more research.

Development and performance evaluation of calf diarrhea pathogen nucleic acid purification and detection workflow.

Calf diarrhea (scours) is a primary cause of illness and death in young calves. Significant economic losses associated with this disease include morbidity, mortality, and direct cost of treatment. Multiple pathogens are responsible for infectious diarrhea, including, but not limited to, Bovine coronavirus (BCV), bovine Rotavirus A (BRV), and Cryptosporidium spp. Identification and isolation of carrier calves are essential for disease management. Texas Veterinary Medical Diagnostic Laboratory current methods for calf diarrhea pathogen identification include electron microscopy (EM) for BCV and BRV and a direct fluorescent antibody test (DFAT) for organism detection of Cryptosporidium spp. A workflow was developed consisting of an optimized fecal nucleic acid purification and multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) for single tube concurrent detection of BCV, BRV, and Cryptosporidium spp., and an internal control to monitor nucleic acid purification efficacy and PCR reagent functionality. In "spike-in" experiments using serial dilutions of each pathogen, the analytical sensitivity was determined to be <10 TCID(50)/ml for BCV and BRV, and <20 oocysts for Cryptosporidium spp. Analytical specificity was confirmed using Canine and Feline coronavirus, Giardia spp., and noninfected bovine purified nucleic acid. Diagnostic sensitivity was ≥98% for all pathogens when compared with respective traditional methods. The results demonstrate that the newly developed assay can purify and subsequently detect BCV, BRV, and Cryptosporidium spp. concurrently in a single PCR, enabling simplified and streamlined calf diarrhea pathogen identification.

Spatio-temporal epidemiology of Tritrichomonas foetus infection in Texas bulls based on state-wide diagnostic laboratory data.

Texas is the largest cattle producing state and suffers severe economic losses due to abortions caused by the protozoan parasite Tritrichomonas foetus. The objective of this study was to use data from the state-wide diagnostic laboratory system of Texas to investigate the occurrence and spatio-temporal distribution of bovine trichomoniasis (BT) in Texas, and to identify spatial disease clusters within the state. The study population consisted of bulls tested for BT in 2010 by the Texas Veterinary Medical Diagnostic Laboratory system that performs at least 95% of all T. foetus testing in the state. Preputial samples were cultured and diagnosis was made by real-time polymerase chain reaction (PCR). Data on BT was aggregated at the county level with time aggregation of one month. The scan statistics was used to identify spatial disease clusters. The database included 31,202 test results with a proportion of positives of 3.7%. As expected, BT was present throughout Texas. Testing prevalence was highest in the summer (5.5%). The scan statistics identified a spatial cluster in southeastern Texas, which could only partially be explained by cattle herd density. The findings of this study provide baseline data to monitor the success of BT control activities in Texas and aids in generating hypotheses regarding specific risk factors for the disease. The identification of high-risk areas and periods is also essential to improve intervention efforts.

Evaluation of factors associated with the risk of infection with Cryptosporidium parvum in dairy calves.

OBJECTIVE: To identify risk factors associated with Cryptosporidium parvum infection in dairy calves. ANIMALS: 108 case animals and 283 control animals. PROCEDURES: Case animals were calves infected with C. parvum, and controls were infected with Cryptosporidium bovis (n = 67) or calves not infected with Cryptosporidium spp. Fecal samples were tested via the flotation concentration method for Cryptosporidium spp. Samples were genotyped by sequencing of the 18s rRNA gene. Associations between host, management, geographic, and meteorologic factors and Cryptosporidium genotype were assessed. RESULTS: Younger calves and calves housed in a cow barn were more likely to be infected with both genotypes. Herd size and hay bedding were associated with an increased risk of infection with C. parvum, and Jersey breed was a risk factor for C. bovis infection. Compared with a flat surface, a steeper slope was significantly associated with a decreased likelihood of infection with both genotypes, and precipitation influenced the risk of C. parvum infection only. CONCLUSIONS AND CLINICAL RELEVANCE: Risk factors for calf infection with C. parvum differed from those for infection with C. bovis. Results may be useful to help design measures that reduce animal exposure and decrease public health risk and economic losses associated with C. parvum infection in cattle.