Role of Plasmids in Co-Selection of Antimicrobial Resistances Among Escherichia coli Isolated from Pigs.

Co-selection is thought to occur when resistance genes are located on the same mobile genetic element. However, this mechanism is currently poorly understood. In this study, complete circular plasmids from swine-derived Escherichia coli were sequenced with short and long reads to confirm that resistance genes involved in co-resistance were co-transferred by the same plasmid. Conjugative transfer tests were performed, and multiple resistance genes were transmitted. The genes possessed by the donor, transconjugant, and plasmid of the donor were highly similar. In addition, the sequences of the plasmid of the donor and the plasmid of the transconjugant were almost identical. Resistance genes associated with statistically significant combinations of antimicrobial use and resistance were co-transmitted by the same plasmid. These results suggest that resistance genes may be involved in co-selection by their transfer between bacteria on the same plasmid.

Evaluation of basilar artery and cerebrospinal fluid dynamics using phase-contrast MRI: Comparison between mannitol and isotonic saline solution.

Increased intracranial pressure (ICP) can cause irreversible pathological changes in the canine brain and can be life-threatening, so prompt diagnosis and therapeutic responses are warranted. The purposes of this prospective experimental study were to evaluate phase-contrast MRI (PC-MRI) as a non-invasive method for quantifying cerebrospinal fluid (CSF) and basilar artery flow, and to assess effects of intravenous administration of hypertonic fluid. A PC-MRI scan was acquired for six healthy Beagle dogs at the level of the mesencephalic aqueduct. Either 1.0 g/kg mannitol or isotonic saline solution was administered intravenously for 15 min each at a matched dose volume of 5 mL/kg. Basilar artery and CSF flow rates were measured and their values compared between mannitol and isotonic saline solution groups before administration, and subsequently every 15 min for 2 h post-administration. The CSF dynamics were further assessed by measuring repeat flow from the caudal to rostral direction and the rostral to caudal direction as the number of waves. No significant difference was observed in basilar or and CSF flow velocity between the two groups (P > .05). However, administration of isotonic saline solution tended to increase basilar artery velocity slightly over time, while CSF velocity remained unchanged. In the mannitol group, CSF wave forms tended to be reduced at 60 and 75 min (P > .05). Findings from this preliminary study indicated that it is feasible to measure the dynamics of CSF and basilar artery flow by PC-MRI, but no flow differences could be detected for mannitol versus isotonic saline administration.

Knowledge, attitudes, and practices (KAP) related to brucellosis and factors affecting knowledge sharing on animal diseases: a cross-sectional survey in the dry zone of Sri Lanka.

Farmers' lack of knowledge is assumed to have affected the presence of brucellosis in Sri Lanka for decades. This study, carried out in the Ampara district in the dry zone of Sri Lanka, revealed that there is a significant knowledge gap for brucellosis compared to foot and mouth disease (FMD) (p < 0.001). Only 8.3% of farmers knew that brucellosis causes cattle abortions. Only 2.6% knew that it is zoonotic. The difference in knowledge of the symptoms and transmission of brucellosis and FMD was significant (p < 0.001). Farmers' attitudes and practices related to the spread of the disease were poor. Farmers' education and spoken language had a negative influence on knowledge. Young people and those with strong social relationships were efficient in knowledge sharing. It can be concluded that brucellosis knowledge, attitudes, and practices are poor; thus, there is a need for more attention in disease control policymaking. Backward farmer groups should be the focus in animal health extension programs.

Low-frequency transmission and persistence of antimicrobial-resistant bacteria and genes from livestock to agricultural soil and crops through compost application.

Livestock excrement is composted and applied to agricultural soils. If composts contain antimicrobial-resistant bacteria (ARB), they may spread to the soil and contaminate cultivated crops. Therefore, we investigated the degree of transmission of ARB and related antimicrobial resistance genes (ARGs) and, as well as clonal transmission of ARB from livestock to soil and crops through composting. This study was conducted at Rakuno Gakuen University farm in Hokkaido, Japan. Samples of cattle feces, solid and liquid composts, agricultural soil, and crops were collected. The abundance of Escherichia coli, coliforms, ß-lactam-resistant E. coli, and ß-lactam-resistant coliforms, as well as the copy numbers of ARG (specifically the bla gene related to ß-lactam-resistant bacteria), were assessed using qPCR through colony counts on CHROMagar ECC with or without ampicillin, respectively, 160 days after compost application. After the application of the compost to the soil, there was an initial increase in E. coli and coliform numbers, followed by a subsequent decrease over time. This trend was also observed in the copy numbers of the bla gene. In the soil, 5.0 CFU g-1 E. coli was detected on day 0 (the day post-compost application), and then, E. coli was not quantified on 60 days post-application. Through phylogenetic analysis involving single nucleotide polymorphisms (SNPs) and using whole-genome sequencing, it was discovered that clonal blaCTX-M-positive E. coli and blaTEM-positive Escherichia fergusonii were present in cattle feces, liquid compost, and soil on day 0 as well as 7 days post-application. This showed that livestock-derived ARB were transmitted from compost to soil and persisted for at least 7 days in soil. These findings indicate a potential low-level transmission of livestock-associated bacteria to agricultural soil through composts was observed at low frequency, dissemination was detected. Therefore, decreasing ARB abundance during composting is important for public health.

Patterns of Animal Rabies Prevalence in Northern South Africa between 1998 and 2022.

Rabies is endemic in South Africa and rabies cycles are maintained in both domestic and wildlife species. The significant number of canine rabies cases reported by the World Organization for Animal Health Reference Laboratory for Rabies at Onderstepoort suggests the need for increased research and mass dog vaccinations on specific targeted foci in the country. This study aimed to investigate the spatiotemporal distribution of animal rabies cases from 1998 to 2017 in northern South Africa and environmental factors associated with highly enzootic municipalities. A descriptive analysis was used to investigate temporal patterns. The Getis-Ord Gi statistical tool was used to exhibit low and high clusters. Logistic regression was used to examine the association between the predictor variables and highly enzootic municipalities. A total of 9580 specimens were submitted for rabies diagnosis between 1998 and 2022. The highest positive case rates were from companion animals (1733 cases, 59.71%), followed by livestock (635 cases, 21.88%) and wildlife (621 cases, 21.39%). Rabies cases were reported throughout the year, with the majority occurring in the mid-dry season. Hot spots were frequently in the northern and eastern parts of Limpopo and Mpumalanga. Thicket bush and grassland were associated with rabies between 1998 and 2002. However, between 2008 and 2012, cultivated commercial crops and waterbodies were associated with rabies occurrence. In the last period, plantations and woodlands were associated with animal rabies. Of the total number of municipalities, five consistently and repeatedly had the highest rabies prevalence rates. These findings suggest that authorities should prioritize resources for those municipalities for rabies elimination and management.

Association of farmers' knowledge, attitude and practices with bovine brucellosis seroprevalence in Myanmar.

OBJECTIVE: This study aimed to identify the relationship between bovine brucellosis prevalence, farmers' knowledge, attitude, practice (KAP), and social factors on migratory draft cattle and smallholder dairy farms in the central dry zone of Myanmar. METHODS: This cross-sectional study was conducted on 54 migratory and 38 dairy cattle farms between August 2020 and February 2021. A structured questionnaire was used to identify farmers' behaviors. Bulk milk was sampled and tested using indirect enzyme-linked immunosorbent assay (I-ELISA). STATA 17 was used for all the analyses. RESULTS: Migratory cattle farms had a higher farm level brucellosis prevalence (14.8%) than dairy farms (2.6%; χ2 = 3.75; df = 1; p = 0.05). Only 2.8% of the farmers had knowledge about brucellosis, while 39.1% and 41.6% had attitudes and farm practices with respect to brucellosis, respectively in the study area. Socio-economic attribute of training in animal husbandry (p<0.01), raising system (p<0.01), practice of separating the aborted cow (p<0.01) were negatively associated to brucellosis. The overall farm level brucellosis prevalence was strongly associated with cattle herd size (p = 0.01), free movement grazing practices (p<0.01), practice of self-removal of placental debris without using personal protective equipment (p<0.01) and farmers' attitudes towards eating cow placenta debris (p<0.01). CONCLUSION: Farmers had little knowledge of brucellosis. Attitudes and practices differed significantly between migratory and dairy farmers. Training and extension programs are necessary to make farmers aware of their KAP situation since livestock migration and the custom of eating cow placental debris contribute to the spread of brucellosis. Persistent efforts are required to reduce the adverse effects of brucellosis. Therefore, the study suggests that a feasible control intervention and public awareness campaigns need to be conducted regarding methods of preventing human exposure to brucellosis.

Evaluation of Immune Status of Pigs against Classical Swine Fever for Three Years after the Initiation of Vaccination in Gifu Prefecture, Japan.

In 2018, classical swine fever (CSF) reemerged in Gifu Prefecture, Japan, after 26 years of absence, and vaccination of domestic pigs using a live attenuated vaccine was initiated in 2019. Because the vaccine efficacy in piglets is influenced by the maternal antibody levels, vaccination should be administered at the optimal age by assuming the antibody level in sows. In this study, the shift in the antibody titer distribution in sows due to the initiation of vaccination to naïve herds and its influence on the vaccine-induced immunity rate in fattening pigs were investigated for 3 years. The results indicated that higher antibody titers were induced in first-generation sows after vaccine initiation because they were immunologically naïve, but the distribution of antibody titers shifted to lower levels along with their replacement with second-generation sows. The average vaccination age of fattening pigs became earlier year by year, and the vaccine-induced antibody rate was almost ≥80%. Based on the estimation of the optimal age for vaccination, it was found that vaccination at a younger age may reduce the risk of CSF infection. Taken together, the risk of CSF outbreaks can be reduced by administering vaccines at the optimal age based on the sequential monitoring of the sow's immune status.

Metabolites and physical scores as possible predictors for postpartum culling in dairy cows.

The purpose of the study was to explore the associations of serum non-esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHBA) concentrations with the body condition score (BCS) and rumen fill score (RFS) in order to predict the risk of postpartum culling, and to further investigate effective monitoring stages during the dry period. From October 2012 to March 2014, clinically healthy Holstein heifers and cows were sampled once before calving, and the occurrence of culling within 60 days in milk (DIM) was investigated. The discriminatory ability of each parameter was evaluated using receiver operating characteristic (ROC) analysis. Of the 720 cows sampled between 14 and 2 days before the actual day of calving in the study, 42 cull cows (mean DIM ± SE: 22.0 ± 2.6) were confirmed. The areas under the curve (AUC) of the ROC for predicting culling using serum NEFA concentrations were 0.6 and 0.7 at 14 to 2 and 7 to 2 days before calving, respectively. The AUC for the RFS was 0.7 for both periods, indicating the same diagnostic level as the serum NEFA concentration. Both the serum NEFA concentration and RFS were possible predictors in cows with ≥ 2 parities, but not in cows with 0-1 parity sampled even at 7 to 2 days before calving. The serum BHBA concentration and BCS were not suitable predictors of culling for any period or parity. These results indicate that RFS has a discriminatory ability comparable to the serum NEFA concentration for predicting culling within 60 DIM.

Evaluating the impact of application of anaerobic bacterial fermentation enhancer on digital dermatitis.

The control of digital dermatitis (DD) among cattle is crucial; however, effective and environmentally-sound control measures have yet to be identified. From the monitoring data of DD which were recorded during regular hoof trimmings in a farm in Hokkaido, Japan, we detected a decrease in the DD prevalence in a herd where an anaerobic bacterial fermentation enhancer (ABFE) was distributed. The possible effect of ABFE was analyzed using a retrospective repeated cross-sectional design. The prevalence of DD decreased over time in the ABFE-distributed group. Furthermore, a selected regression model indicated the time-dependent enhancement of the decreasing trend. While potential coincidental factors may influence, this study provides a basis for further research on the preventive effect of ABFE against DD.

The final stages of dog rabies elimination from Japan.

Rabies is a lethal zoonotic disease mainly transmitted to humans by dog bites. The purpose of this study was to assess the efficacy of rabies control policies in Japan, which resulted in the elimination of the disease from the country in 1957. Using historical records from the Kanto region (Chiba, Kanagawa, Saitama and Tokyo Prefectures) between 1947 and 1956 where the final canine cases were recorded, we undertook a descriptive epidemiological study, applying spatio-temporal scan statistics using SaTScan and estimating the effective reproduction number (Rt ) for the clusters and each prefecture using the growth rates. There were 1,567 dog rabies and 161 human rabies cases recorded during this period. Vaccination coverage in registered dogs was over 70% after 1951, with much lower coverage in free-roaming and unregistered dogs. Eight clusters of dog rabies cases were identified: the first appeared in 1947 in Tokyo and was linked to three further clusters in peripheral prefectures between 1947 and 1951. Three more clusters occurred in Tokyo again between 1952 and 1954, and the last cluster was in Tokyo and Kanagawa between 1955 and 1956. Rt in the first cluster was 1.68, and Rt values in the others ranged between 1.18 and 1.86, with an exception of 4.05 in the smallest cluster in Tokyo in 1952 (10 cases). The moving average of Rt coincided with the clusters. As dog vaccination and dog management progressed, and the number of dog rabies cases declined, the moving average of Rt declined to below 1. Delays in the implementation of dog management policies in Kanagawa may have prolonged this last outbreak. These results demonstrate the effectiveness of coordinated control policy involving dog vaccination and management of free-roaming dog populations for rabies elimination.

Hazard identification and exposure assessment for bacterial risk assessment of informally marketed milk in Abidjan, Côte d'Ivoire.

BACKGROUND: Animal-source foods are important causes of food-borne illness, and milk and dairy products can contain pathogenic microorganisms. OBJECTIVE: We conducted a stochastic assessment of the risk of ingesting milk contaminated with specific microbial pathogens (Escherichia coli, Staphylococcus aureus, and Enterococcus spp.) in Abidjan, Côte d'Ivoire. METHODS: We carried out structured interviews and focus group discussions with farmers (n = 15), vendors (n = 17), and consumers (n = 188) to characterize dairy production systems and milk consumption behavior. Microbiological sampling was conducted at different points between milking and sale. A risk model was developed, and the risk of consuming contaminated raw milk was estimated by Monte Carlo simulation. RESULTS: The investigation into local raw milk consumption patterns showed that the proportion of raw milk consumption was 51.6% in people who consume milk. The probability of ingestion of marketed raw milk that failed to meet standards for this group of bacteria was 29.9% and about 652 consumers per day were estimated to ingest contaminated milk. Microbiological tests from the farm showed that 7.2% of samples taken from milkers' hands, 4.4% of water samples (water used to rinse milk containers or milking utensils (calabash, plastic bottle, filters, buckets), 4.4% of environmental samples (air pollution), 13.2% of samples from milking utensils, and 4.9% of samples from cows' udders were contaminated with one or more of these pathogens. About 624.6 L of marketed raw milk would need to be discarded per day if discarding milk was chosen as the option of risk reduction. The destruction of this milk would result in a potential loss of Euro623.9 per day for all producers. CONCLUSIONS: The risk of human illness from consumption of raw milk could be mitigated by raising awareness about heat treatment of milk and good hygiene practices in the dairy chain.

Cardiovascular effects of intravenous pimobendan in dogs with acute respiratory acidosis.

OBJECTIVE: Acidosis decreases myocardial contractile and myofibrillar responsiveness by reducing the calcium sensitivity of contractile proteins, which could reduce the effectiveness of pimobendan. We aimed to assess the cardiovascular effects of pimobendan in dogs subjected to acute respiratory acidosis. DESIGN: Randomized crossover study with a 2-week washout period. SETTING: University Laboratory. ANIMALS: Six healthy research Beagle dogs. INTERVENTIONS: Anesthetized dogs were administered 2 doses of IV pimobendan during conditions of eucapnia (Paco2 35-40 mm Hg) and hypercapnia (Paco2 90-110 mm Hg). Eucapnia was maintained by positive pressure ventilation and hypercapnia was induced by adding exogenous CO2 to the anesthesia circuit. Heart rate (HR), systemic arterial blood pressure, cardiac output (CO), systemic and pulmonary vascular resistance (SVR and PVR, respectively), and pulmonary arterial pressure (PAP) were measured at baseline and 60 min after administering 0.125 mg/kg (low) and 0.25 mg/kg (high) pimobendan intravenously. Blood gas and biochemical analyses were performed at baseline and at the end of the experiment. MEASUREMENTS AND MAIN RESULTS: The median baseline blood pH was 7.41 (range: 7.33-7.45) and 7.03 (range: 6.98-7.09) under conditions of eucapnia and hypercapnia, respectively. The serum concentrations of epinephrine and norepinephrine and the HR, CO, and PAP were higher, and SVR was lower at baseline in hypercapnic dogs. Pimobendan dose-dependently increased CO in eucapnia (baseline: 3.6 ± 0.2 L/kg/m2 [mean ± SE], low: 5.0 ± 0.4 L/kg/m2 , high: 5.8 ± 0.5 L/kg/m2 , P < 0.001) and hypercapnia (baseline: 4.9 ± 0.5 L/kg/m2 , low: 5.8 ± 0.5 L/kg/m2 , high: 6.2 ± 0.5 L/kg/m2 , P < 0.001), and increased HR and decreased SVR and PVR under both conditions (P < 0.001). In hypercapnia, the degree of increase or decrease of these cardiovascular measurements (except for PAP) by pimobendan was less than that in the eucapnic dogs. CONCLUSIONS: Pimobendan maintains function as an inodilator in anesthetized dogs with induced respiratory acidosis.

Use of a Participatory Method for Community-Based Brucellosis Control Design in Agro-Pastoral Areas in Tanzania.

Brucellosis is widespread in both humans and livestock in many developing countries. The authors have performed a series of epidemiological studies on brucellosis in agro-pastoral areas in Tanzania since 2015, with the aim of the disease control. Previously, the potential of a community-based brucellosis control initiative, which mainly consisted of the sale of cattle with experience of abortion and vaccinating calves, was assessed as being effective and acceptable based on a quantitative approach. This study was conducted to investigate the feasibility of community-based brucellosis control program using participatory rural appraisals (PRAs) and key-informant interviews. Four PRAs were performed together with livestock farmers and livestock and medical officers in 2017. In the PRAs, qualitative information related to risky behaviors for human infection, human brucellosis symptoms, willingness to sell cattle with experience of abortion, and willingness to pay for calf vaccination were collected, and a holistic approach for a community-based disease control project was planned. All of the communities were willing to implement disease control measures. To avoid human infection, education, especially for children, was proposed to change risky behaviors. The findings of this study showed that community-based disease control measures are promising.

Estimation of economic loss by carcass weight reduction of Japanese dairy cows due to infection with bovine leukemia virus.

Bovine leukemia virus (BLV) infection is endemic in Japanese dairy farms. To promote the participation of farmers in BLV infection control in Japan, it is important to provide estimates of the economic losses caused by this infection. We hypothesized that decreased immune function due to BLV infection would increase visceral abnormalities, in turn reducing carcass weight. We employed mediation analysis to estimate the annual economic loss due to carcass weight reduction caused by BLV infection. Culled Holstein cows from 12 commercial dairy farms in the Nemuro and Kushiro regions of Hokkaido, Japan, were traced. Information on age and the last delivery day were collected. A non-infected culled cow was defined as a cow from which BLV provirus was not detected. A high-proviral-load (H-PVL) cow was defined as a cow whose PVL titer was above 2465 copies/50 ng DNA or 56,765 copies/105 cells. A BLV-infected cow with PVL titer lower than the thresholds was categorized as low-proviral load (L-PVL). Post-mortem examination results for culled cows were collected from a meat inspection center. The hypothesis was tested by three models, using data from 222 culled dairy cows. Model 1, a generalized linear mixed-effects model, selected carcass weight as an outcome variable, BLV status and the potential confounders (lactation stage and age) as explanatory variables, and herd as a random effect. Model 2 additionally included the number of abnormal findings in the post-mortem examination (AFPE) as an explanatory variable. Model 3 applied a Bayesian generalized linear mixed model, which employed a mediator separately modeled for AFPE, to estimate the amount of direct, indirect, and total carcass weight loss with adjustment for known confounding factors. Compared to the mean carcass weight for the non-infected culled cows, the carcass weight for H-PVL culled cows was significantly decreased by 30.4 kg on average. For each increase of one in the number of AFPE, the mean carcass weight was decreased by 8.6 kg. Only the indirect effect of BLV H-PVL status on carcass weight loss through AFPE was significant, accounting for 21.6 % of the total effect on carcass weight reduction. In 2017, 73,650 culled dairy cows were slaughtered in Hokkaido, and the economic loss due to carcass weight loss caused by BLV infection that year was estimated to be US $1,391,649. In summary, unlike L-PVL cows, H-PVL status was associated with carcass weight reduction, which was partially mediated by an increase in the number of visceral abnormalities.

The impact of coronavirus disease 2019 (COVID-19) in Japanese companion animal clinics.

The coronavirus disease 2019 (COVID-19) pandemic has had a tremendous impact on people's lives throughout the world. A cross-sectional study was conducted to clarify the influence of COVID-19 on Japanese companion animal clinics. A self-administered electronic questionnaire regarding the incidence of COVID-19, hygiene management, the influence on clinical service and employment, and mental stress of staff was conducted for workers in animal clinics between 1 May and 10 June 2021. Questions concerning the hygiene management before the occurrence of COVID-19, under the first state of emergency, and at the time of the survey were asked using the Likert scale. Kessler 6 (K6) was used as an indicator of mental distress. In total, 430 individuals responded. Of these, 4.9% experienced COVID-19 infection in staff. Hygiene management was strengthened, but no particular practice was statistically associated with the occurrence of COVID-19 cases in staff. In Nakaya's variation of Scheffe's paired comparison, the highest prioritized goal in the clinics was the prevention of nosocomial infection with COVID-19. The prevalence of serious mental illness (K6 >=13) was 11.1% (95% confidence interval: 6.3-18.6%). Multivariable negative binomial regression found four risk factors for psychological distress: veterinary nurse (P=0.016 with veterinarians and P<0.01 with other staff), female (P=0.004), fear of infection at work (P<0.001), and stress by refraining from going out (P<0.001). Directors of clinics are recommended to take care of female veterinary nurses for distress.

Spatio-temporal epidemiology of animal and human rabies in northern South Africa between 1998 and 2017.

BACKGROUND: Rabies is a fatal zoonotic disease that is maintained in domestic dogs and wildlife populations in the Republic of South Africa. A retrospective study was conducted to improve understanding of the dynamics of rabies in humans, domestic dogs, and wildlife species, in relation to the ecology for three northern provinces of South Africa (Limpopo, Mpumalanga, and North-West) between 1998 and 2017. METHODS: A descriptive epidemiology study was conducted for human and animal rabies. Dog rabies cases were analyzed using spatio-temporal scan statistics. The reproductive number (Rt) was estimated for the identified disease clusters. A phylogenetic tree was constructed based on the genome sequences of rabies viruses isolated from dogs, jackals, and an African civet, and Bayesian evolutionary analysis using a strict time clock model. Several ecological and socio-economic variables associated with dog rabies were modeled using univariate analyses with zero-inflated negative binomial regression and multivariable spatial analyses using the integrated nested Laplace approximation for two time periods: 1998-2002 and 2008-2012. RESULTS: Human rabies cases increased in 2006 following an increase in dog rabies cases; however, the human cases declined in the next year while dog rabies cases fluctuated. Ten disease clusters of dog rabies were identified, and utilizing the phylogenetic tree, the dynamics of animal rabies over 20 years was elucidated. In 2006, a virus strain that re-emerged in eastern Limpopo Province caused the large and persistent dog rabies outbreaks in Limpopo and Mpumalanga Provinces. Several clusters included a rabies virus variant maintained in jackals in Limpopo Province, and the other variant in dogs widely distributed. The widely distributed variant maintained in jackal populations in North-West Province caused an outbreak in dogs in 2014. The Rt was high when the disease clusters were associated with either multiple virus strains or multiple animal species. High-risk areas included Limpopo and Mpumalanga Provinces characterized by woodlands and high temperatures and precipitation. CONCLUSION: Canine rabies was maintained mainly in dog populations but was also associated with jackal species. Rural communities in Limpopo and Mpumalanga Provinces were at high risk of canine rabies originating from dogs.

Decreasing the abundance of tetracycline-resistant Escherichia coli in pig feces during nursery using flavophospholipol as a pig feed additive.

Tetracyclines (TCs) are widely used for livestock, and the high prevalence of TC-resistant Escherichia coli in livestock has become a serious concern worldwide. In Japan, the National Action Plan on Antimicrobial Resistance in 2016 aimed to reduce the TC resistance rate in E. coli derived from livestock. Flavophospholipol (FPL), used as a feed additive, has an inhibitory effect on the spread of plasmid-mediated antimicrobial resistance. The number of TC-resistant E. coli was determined in pigs administered TCs and/or FPL to clarify the effect of FPL on reducing the number of TC-resistant E. coli in pigs. TC-resistant E. coli and their plasmids were then analyzed. The pigs were divided into four groups: control, doxycycline (DOXY; a TC), FPL, and a DOXY-FPL combination. Their feces were collected from the nursing period to the day before being transported to the slaughterhouse, followed by estimation of TC-resistant E. coli (colony-forming units [CFU]/g). The number of TC-resistant E. coli increased with the use of DOXY, suggesting that DOXY administration provides a selective pressure for TC-resistant E. coli. Supplementation with FPL as a feed additive significantly suppressed the increase in the number of TC-resistant E. coli, especially during the DOXY administration period. Transfer and growth inhibition analyses were performed for TC-resistant isolates. FPL inhibited the conjugational transfer and growth of a few TC-resistant E. coli isolates. These results suggest that FPL is effective against the spread of TC-resistant E. coli.

Herd prevalence of bovine brucellosis and analysis of risk factors in cattle in urban and peri-urban areas of the Kampala economic zone, Uganda.

BACKGROUND: Human brucellosis has been found to be prevalent in the urban areas of Kampala, the capital city of Uganda. A cross-sectional study was designed to generate precise information on the prevalence of brucellosis in cattle and risk factors for the disease in its urban and peri-urban dairy farming systems. RESULTS: The adjusted herd prevalence of brucellosis was 6.5% (11/177, 95% CI: 3.6%-10.0%) and the adjusted individual animal prevalence was 5.0% (21/423, 95% CI: 2.7%-9.3%) based on diagnosis using commercial kits of the competitive enzyme-linked immunosorbent assay (CELISA) for Brucella abortus antibodies. Mean within-herd prevalence was found to be 25.9% (95% CI: 9.7%-53.1%) and brucellosis prevalence in an infected herd ranged from 9.1% to 50%. A risk factor could not be identified at the animal level but two risk factors were identified at the herd level: large herd size and history of abortion. The mean number of milking cows in a free-grazing herd (5.0) was significantly larger than a herd with a movement restricted (1.7, p < 0.001). CONCLUSIONS: Vaccination should be targeted at commercial large-scale farms with free-grazing farming to control brucellosis in cattle in and around Kampala city.

Spatial epidemiology of hospital-diagnosed brucellosis in Kampala, Uganda.

BACKGROUND: A retrospective case-control study was undertaken to examine the spatial risk factors for human brucellosis in Kampala, Uganda. METHODS: Information on age, sex and month of diagnosis was derived from records from plate agglutination tests undertaken at Mulago Hospital, Kampala. Information on Parishes (LC2s) where patients reside was sourced from the outpatient registration book. In-patient fracture cases were selected for use as controls using 1:1 matching based on the age, sex and month of diagnosis. The locations of cases and controls were obtained by calculating Cartesian coordinates of the centroids of Parish level (LC2) polygons and a spatial scan statistic was applied to test for disease clustering. Parishes were classified according to the level of urbanization as urban, peri-urban or rural. RESULTS: Significantly more females than males were found to show sero-positivity for brucellosis when compared with the sex ratio of total outpatients, in addition female brucellosis patients were found to be significantly older than the male patients. Spatial clustering of brucellosis cases was observed including around Mulago Hospital (radius = 6.8 km, p = 0.001). The influence of proximity to the hospital that was observed for brucellosis cases was not significantly different from that observed in the controls. The disease cluster was confounded by the different catchment areas between cases and controls. The level of urbanization was not associated with the incidence of brucellosis but living in a slum area was a significant risk factor among urban dwellers (odds ratio 1.97, 95% CI: 1.10-3.61). CONCLUSIONS: Being female was observed to be a risk factor for brucellosis sero-positvity and among urban dwellers, living in slum areas was also a risk factor although the overall risk was not different among urban, peri-urban and rural areas of the Kampala economic zone.

Decreased colistin resistance and mcr-1 prevalence in pig-derived Escherichia coli in Japan after banning colistin as a feed additive.

BACKGROUND: Antimicrobial resistance to colistin, a widely used feed additive for farm animals across the world, has raised public health concern in recent years. Since July 2018, its use as feed additive has been banned in Japan to reduce the spread of plasmid-based mobilized colistin resistance (mcr) genes and the subsequent development of colistin-resistant bacteria. Evaluating the effects of these measures is required. METHODS: We evaluated the effects of colistin use, as a feed additive, on colistin resistance in pigs (n=5) from birth to finishing in the farm. Moreover, to evaluate changes in colistin resistance and mcr gene prevalence in response to colistin withdrawal, E. coli samples derived from pig faeces sourced from the fields of three geographically distinct farms were characterized before and after the withdrawal of colistin as a feed additive. RESULTS: Colistin-resistant Escherichia coli in pigs (n=5) increased during the colistin administration period and decreased immediately after its end. In three fields, the colistin resistance rate and prevalence of mcr-1 decreased immediately and significantly after the ban. However, colistin-resistant and mcr-1-positive E. coli were still detected in all three farm fields 12 months after the ban on colistin use. CONCLUSION: Agricultural colistin use caused selective pressure that contributed to widespread mcr dissemination in Japan. Colistin resistance and the presence of mcr genes should be continuously monitored in food-producing animals.