Effect of Mycoplasma bovis on expression of inflammatory cytokines and matrix metalloproteinases mRNA in bovine synovial cells.

Mycoplasma bovis causes chronic arthritis in calves. Mycoplasma arthritis shows severe inflammatory reactions in joints that is commonly treated with antibiotics and results in significant economic losses in the calf industry. A previous study showed that inflammatory cytokines and matrix metalloproteinases (MMPs) produced by synovial cells promote progression of the pathophysiology of bacterial arthritis. However, the mechanism underlying the pathogenesis of bovine Mycoplasma arthritis has not been fully clarified. In this study, we examined the immunologic response of bovine synovial tissue to M. bovis. We observed significant increases in expression of interleukin (IL)-1ß, IL-6, IL-8, MMP-1, and MMP-3 mRNA in synovial tissue from Mycoplasma arthritis calves compared with tissues from normal calves. Expression of IL-6, IL-8, and MMP-1 mRNA was also induced in cultured synovial cells stimulated with M. bovis, but not expression of IL-1ß and MMP-3 mRNA. In contrast, the culture supernatant of peripheral blood mononuclear cells stimulated with M. bovis induced marked increases in the expression of IL-1ß, IL-6, IL-8, MMP-1, and MMP-3 mRNA in synovial cells. Our results indicate that inflammatory cytokines and MMPs produced by synovial cells play a key role in the pathogenesis of Mycoplasma arthritis. We suggest that interactions between synovial cells and mononuclear cells in the presence of M. bovis induce expression of these cytokines and MMPs in synovial cells, resulting in severe inflammatory reactions in the joints.

Prevalence and risk factors of Mycoplasma bovis infection in dairy farms in northern Japan.

Mycoplasma mastitis is a dairy herd health problem with growing concern in Japan. To complement the lack of epidemiological knowledge of the disease, we conducted estimation of herd-level prevalence and risk factor analysis for Mycoplasma bovis (M. bovis) mastitis using data collected from Tokachi region, one of the nation's largest milk producing area, in Hokkaido Prefecture in 2015. The herd-level prevalence was estimated at 3.8% (95% confidence interval (CI): 2.6%, 5.4%) across the region with municipality-specific prevalence ranging from 0% to 14.3%. In this study, identified risk factors for the presence of with M. bovis infection on farms were corporation-type farms and purchased cattle, after controlling for the herd size. Corporation-type farms may reflect higher frequencies of moving cows and fomites to and from other farms, which increases the risk of pathogen introduction. Purchased cattle was considered as one of the major pathways of the disease incursion, and this finding highlighted the importance of more stringent separation or quarantine protocols when introducing cattle from outside in Tokachi region. Due to the limited information available in this study and inherent nature of the study designs, these results should be interpreted with caution and further research is needed.

Matched case-control study of the influence of inland waters surrounding poultry farms on avian influenza outbreaks in Japan.

To successfully control highly pathogenic avian influenza (HPAI), understanding the risk factors related to the incursion of the virus into poultry farms is essential. In this study, we focused on the presence of inland waters surrounding poultry farms as a potential risk factor of incursion of the virus. To evaluate the influence of inland waters surrounding poultry farms on HPAI outbreaks in Japan, a simple matched case-control study was conducted. The results of the conditional regression analyses indicated that the number of farms with neighbouring inland waters was significantly high among the affected farms during the 2016-2017 outbreak period. These results provide good grounds for strengthening biosecurity management at farms located near inland waters.

Mathematical modeling of porcine epidemic diarrhea virus dynamics within a farrow-to-finish swine farm to investigate the effects of control measures.

Porcine epidemic diarrhea (PED) is a highly contagious enteric disease in swine that can cause devastating economic damage to pig producers. Japan was severely affected by PED epidemics from 2013 through 2015, with over 1000 farms were affected during this period. Although many studies have unraveled pathological and molecular characteristics of PED virus (PEDV), the mechanism for within-farm spread is largely unknown. Here, we constructed a deterministic compartmental model to quantitatively describe the infection dynamics in a farm setting and to investigate effective control measures. The model consisted of three separate houses and four swine populations framed in a standard commercial farrow-to-finish swine operation in Japan, with a special focus on the role of indirect transmission via the on-farm environment contaminated by feces of infected pigs. Some model parameters were estimated using the Japanese empirical outbreak data. Model outputs over a 90-day period showed that the number of infected sows in a dry sow house peaked within a week after disease introduction, and PEDV was retained on the farm. In the farrowing house, the number of infected sows also peaked within a week followed by a tentative disappearance of infected pigs, whereas the number of infected piglets in the farrowing house spiked and the number of neonatal deaths increased rapidly until around Day 21 followed by a continued increase at a slower pace. In the fattening house, the number of infected pigs reached an equilibrium at around Day 25 and approximately 15% of pigs continued to be infected thereafter. Herd size was not found to have a significant influence on the overall trend in our model output. Among the control measures examined using our model, the application of effective vaccination was considered to be the most promising. Activities such as performing more thorough cleanings to remove residual virus or implementing stricter on-farm biosecurity to cease between-house virus transfer demonstrated little effectiveness for the control of PED. These results may imply that preventing PEDV incursion on a swine farm by elevated farm biosecurity measures is critically important, and that further research is necessary to prepare for future outbreaks, particularly research related to development of an effective vaccine that can prevent infection, and/or reduction of piglet mortality.

Evaluation of fecal shedding and antibody response in dairy cattle infected with paratuberculosis using national surveillance data in Japan.

Paratuberculosis or Johne's disease (JD), is a chronic infectious disease causing intractable diarrhea in cattle, which leads to less productivity, such as decreased milk yield, and lower daily weight gain. As a control measure against JD in cattle, national serological surveillance has been conducted in Japan since 1998. To conduct modeling studies that are useful to evaluate the effectiveness of control measures against JD, reliable parameter values, such as length of time from infection to the start of fecal shedding or antibody expression, are especially important. These parameters in the Japanese cattle population are assumed to be different from those in other countries with a higher prevalence of JD or in experimental infection settings; therefore, they must be estimated for the cattle population in Japan. Data from national surveillance conducted in Tokachi District, Hokkaido Prefecture, were used for this study. Using data from JD diagnostic tests for all cattle in Tokachi District between 1998 and 2014, all testing histories for infected animals were estimated as the number of tested cattle and positive cattle at each age of month for both fecal and antibody tests. A deterministic mathematical model for JD development, from infection to fecal shedding and antibody expression in infected cattle, was constructed to obtain the probability of testing positive when applied to both fecal and antibody tests at a given age. Likelihood was obtained from these estimated test results and best values for parameters were obtained using the Markov Chain Monte-Carlo method. Fifty-five percent of infected cattle were projected to have a transient shedding period, which was estimated to start 12 months after infection and last for 4 months. Persistent shedding was projected to occur in all infected cattle, and estimated to begin 7-84 months from infection. Following persistent shedding, antibody expression was estimated to start 7 months later. These values are useful for developing models to evaluate the status of JD infection and the effectiveness of control measures in the Japanese cattle population.

Meteorological factors affecting seroconversion of Akabane disease in sentinel calves in the subtropical Okinawa Islands of Japan.

Akabane virus, the pathogen-causing Akabane disease, is an arthropod-borne virus (arbovirus) transmitted by the Culicoides biting midge. A nationwide serological surveillance program for bovine arboviral diseases, including Akabane disease, has been established in Japan to monitor the circulation of arboviruses by targeting sentinel calves. Okinawa, which is located in the southwestern-most region of Japan, is a high-risk area for incursion of arboviruses. The aim of the present study was to identify the meteorological factors related to farm-level seroconversion of Akabane virus by analyzing the serological surveillance data for sentinel calves collected in Okinawa between 2007 and 2015. Rainfall in winter, spring, and autumn was positively associated with seroconversion. Adequate rainfall seems to keep the soil in a suitably moist state for growth and survival of biting midges. Maximum temperature in winter was also positively associated with seroconversion in sentinel calves. The warmer temperatures in winter may provide conditions suitable for shortening the larval development cycle and increase production of adult midges. Our findings indicate that meteorological factors such as temperature and rainfall may be important factors that produce circumstances conducive to effective transmission of the virus between vectors and the host. The results of this study provide a better understanding of the circulation of arboviruses and offer suggestions for developing better surveillance and measures to prevent arboviral disease.

Epidemiological analysis of bovine ephemeral fever in 2012-2013 in the subtropical islands of Japan.

BACKGROUND: Bovine ephemeral fever (BEF) is a febrile disease of cattle that is transmitted by arthropod vectors such as mosquitoes and Culicoides biting midges. An outbreak of BEF recently occurred in Ishigaki Island and surrounding islands that are located southwest of Japan. In this study, an epidemiological analysis was conducted to understand the temporal and spatial characteristics of the outbreak. Factors associated with the disease spread within Ishigaki Island were investigated by hierarchical Bayesian models. The possibility of between-island transmission by windborne vectors and transmission by long-distance migration of infected vectors were examined using atmospheric dispersion models. RESULTS: In September 2012, the first case of the disease was detected in the western part of Ishigaki Island. In 1 month, it had rapidly spread to the southern part of the island and to surrounding islands, and led to 225 suspected cases of BEF during the outbreak. The dispersion model demonstrated the high possibility of between-island transmission by wind. Spatial analysis showed that paddy fields, farmlands, and slope gradients had a significant impact on the 1-km cell-level incidence risk. These factors may have influenced the habitats and movements of the vectors with regard to the spread of BEF. A plausible incursion event of infected vectors from Southeast Asia to Ishigaki Island was estimated to have occurred at the end of August. CONCLUSION: This study revealed that the condition of a terrain and land use significantly influenced disease transmission. These factors are important in assessing favorable environments for related vectors. The results of the dispersion model indicated the likely transmission of the infected vectors by wind on the local scale and on the long-distance scale. These findings would be helpful for developing a surveillance program and developing preventive measures against BEF.

Spatial epidemiological analysis of bovine encephalomyelitis outbreaks caused by Akabane virus infection in western Japan in 2011.

Akabane disease, which is distributed in temperate and tropical regions in the world, is a vector-borne disease of ruminants caused by the Akabane virus, transmitted by Culicoides biting midges. In 2011, outbreaks of Akabane viral encephalomyelitis occurred in the Shimane Prefecture in western Japan. In this study, a spatial epidemiological analysis was conducted to understand environmental factors associated with the spread of Akabane disease. By applying a conditional autoregressive model, the relationship between infection and environmental variables was explored. The results showed that the dominance of farmlands and the presence of infected farms within a 3-km radius had a significant effect on infection. This result implies that land use, which would relate with the vector habitat, and the presence of neighboring infected farms as a source of infection may have influenced the spread of the disease in this region. These findings provide basic insights into the spread of Akabane disease and useful suggestions for developing a surveillance program and preventive measures against the disease.

Cost-effectiveness of diagnostic strategies using quantitative real-time PCR and bacterial culture to identify contagious mastitis cases in large dairy herds.

Diagnostic strategies to detect contagious mastitis caused by Mycoplasma bovis, Staphylococcus aureus, and Streptococcus agalactiae in dairy herds during an outbreak have been minimally studied with regard to cost and diagnostic sensitivity. The objective of this cross-sectional study was to compare the cost-effectiveness of diagnostic strategies for identification of infected cows in two California dairy herds during contagious mastitis outbreaks. M. bovis was investigated in a subset of a herd (n=1210 cows) with an estimated prevalence of 2.8% (95% CI=1.9, 3.7), whereas Staph. aureus and Strep. agalactiae were studied in a second herd (n=351 cows) with an estimated prevalence of 3.4% (95% CI=1.5, 5.3) and 16.8% (95% CI=12.9, 20.7), respectively. Diagnostic strategies involved a combination of testing stages that utilized bacterial culture, quantitative real-time PCR (qPCR), or both. Strategies were applied to individual or pooled samples of 5, 10, 50 or 100 samples. Culture was considered the gold standard for sensitivity estimation of each strategy. The reference strategy was the strategy with the lowest cost per culture-positive cow which for both M. bovis and Strep. agalactiae consisted of 2 stages, culture of samples in pools of 5 followed by culture of individual samples in positive pools with a sensitivity of 73.5% (95% CI: 55.6, 87.1) and 96.6% (95% CI: 27.7, 84.8), respectively. The reference strategy for Staph. aureus consisted of 3 stages, culture of individual samples in pools of 100 (stage 1), culture constituents of those positive from stage 1 in pools of 5 (stage 2), culture constituents of those positive from stage 2 individually (stage 3) which resulted in a sensitivity of 58.3% (95% CI: 88.3, 99.6). The most cost-effective alternative to the reference strategy was whole herd milk culture for all 3 pathogens. QPCR testing was a component of the second most cost-effective alternative for M. bovis and the third most cost-effective alternatives for the 3 pathogens. A stochastic model was used to assess the effect of prevalence or herd size on the cost-effectiveness of diagnostic strategies. In the current study, increasing the prevalence of mastitis did not alter the ranking of strategies by cost-effectiveness. However, larger herds could benefit from testing larger pools such as 50 or 100 samples to improve cost-effectiveness. Several diagnostic strategy options exist to identify contagious mastitis in herds, decisions should be based on cost and sensitivity of the strategies available.