Ryoko Uemura

Comparative studies of the persistence of animal mycoplasmas under different environmental conditions

A comparison of the persistence of mycoplasmas in animals was carried out. When inoculated into liquid media, strains of Mycoplasma bovis, M. arginini, Acholeplasma laidlawii, and A. axanthum persisted for 59-185 days post-inoculation. The survival periods were not significantly influenced by temperature (4, 30, 37 degrees C, and room temperature). The survival periods for M. bovigenitalium, M. gallisepticum, M. bovirhinis, and M. gateae ranged from <7 to 185 days depending on medium components and temperature. Further, it was determined that strains of M. bovigenitalium, M. bovis, M. bovirhinis, M. arginini, and A. laidlawii persisted in a dry paper disc for at most 28, 126, 154, 56 and over >168 days at 4 degrees C, respectively. At 4 degrees C, strains of M. gallisepticum, M. columborale, M. edwardii, M. felis, and M. gateae survived for at most 28, 21, 42, 28, 28 and 70 days, respectively. At 30 degrees C, strains of M. bovis, M. bovirhinis, M. arginini, A. laidlawii, and M. gallisepticum persisted for at most 28, 84, 56, >168 and 14 days, respectively, but strains of M. gallisepticum, M. columborale, M. edwardii, M. felis, M. gateae, and U. diversum did not survive for more than 14 days. In an outdoor environment, strains of M. bovirhinis and A. laidlawii survived for at most 28 and 14 days, respectively. Finally, it was found that 14 isolates of M. gallisepticum persisted for periods similar to those of the reference strains. The results under dry conditions at a variety of temperatures presented contribute to understanding the epizootiology of mycoplasmal infections in the field.

Antimicrobial susceptibilities of Shiga toxin-producing Escherichia coli isolates from pigs with edema disease in Japan.

Fifty‐seven Shiga toxin‐producing Escherichia coli (STEC) strains isolated from pigs with edema disease (ED) from 1997 to 2001 in Japan were examined for antimicrobial susceptibilities. The susceptibilities were compared with those of E. coli ATCC 23546 isolated from pig with ED in the 1950s. Consequently, the isolated STECs showed high susceptibility to peptides and bicozamycin in a way similar to the reference strain. On the other hand, the STECs showed low susceptibility to beta‐lactams, tetracyclines, novobiocin, fosfomycin, trimethoprim, and old quinolones. It became clear that the susceptibilities of the isolated STECs had diminished in regard to antimicrobials.

An assessment of reproductive and lifetime performances of Kagoshima Berkshire gilts and sows.

We investigated the reproductive and lifetime performances of Kagoshima Berkshire gilts and sows. We examined 20 605 parity records of 4419 pigs for 2008 to 2012 on a farrow-to-finish commercial farm. The mean parity (± SD) of all animals was 3.0 ± 2.1. For farrowing performance, the highest numbers of total pigs born and pigs born alive were found in sows with parities 5 and 6 and with parity 3-6, respectively (P < 0.05). Regarding weaning and mating performance, sows with parity 2 had the lowest preweaning mortality (P < 0.05). The longest weaning-to-first-mating interval was found in parity 1 pigs, and the interval decreased as parity increased (P < 0.05). Parities 0 and 1 pigs had the lowest farrowing rate and those with parity 4 had the highest farrowing rate (P < 0.05). The mean parity at culling, total number of pigs born alive in a lifetime, and nonproductive days in a lifetime were 5.5 ± 2.93, 49.2 ± 24.72 pigs, and 132.1 ± 83.34 days, respectively. These animals had a lower litter size and fertility that the F1 crossbred sows mainly used in Japan, but a similar tendency for performance by parity.

Effect of Bicozamycin on the Eradication of Shiga Toxin‐Producing Escherichia coli in Calves

Fifty‐nine calves, aged 11 days to 9 months, from three farms breeding Japanese Black beef cattle in Miyazaki Prefecture, Japan, were examined for the presence of Shiga toxin‐producing Escherichia coli (STEC). A high prevalence of STEC was detected among calves, with 45 (76.3%) animals carrying STEC including different serogroups (O26, O74, O111, O114, O119, O127, O153, O157, and ONT) and toxin types. The number of STEC in the feces was estimated by a combined method involving enumeration of colony‐forming units by a plate‐most‐probable‐number (plate‐MPN) technique and polymerase chain reaction for the detection of Shiga toxin genes. Fecal shedding ranged from 101 to 10s MPN/g feces. To evaluate the safety and efficacy of bicozamycin (BCM: previously named as bicyclomycin) in eradicating STEC, 30 calves carrying STEC with or without diarrhea were examined. Fifteen calves were treated orally with BCM (10 mg/kg/day) once daily for 5 days, and the other 15 were untreated. Twenty‐four hours after the last dose, fecal specimens were collected from both groups to compare the number of coliforms and STEC with those before treatment. BCM‐treated animals had a significantly lower number of coliforms and STEC compared to the untreated calves. The STEC eradication rate was 86.6% (13/15) in the BCM‐treated group, compared to 0% (0/15) in the control group. The corresponding cure rates for diarrhea were 87.5 (7/8) and 0% (0/3), respectively. No adverse reactions were observed in the calves during treatment. It is concluded that BCM is an effective agent for the eradication of STEC in calves with or without diarrhea.

Campylobacter and Salmonella are prevalent in broiler farms in Kyushu, Japan: results of a 2‐year distribution and circulation dynamics audit

To elucidate the distribution and circulation dynamics of Campylobacter and Salmonella in Japanese chicken broiler flocks.

An improved loop-mediated isothermal amplification assay for the detection of Mycoplasma bovis

We improved a loop-mediated isothermal amplification (LAMP) assay permitting sensitive and rapid Mycoplasma bovis detection. A total of 55 bacterial strains were examined in this study, including 33 M. bovis strains, 14 non-M. bovis mycoplasmas and eight non-mycoplasma bacterial strains. M. bovis was successfully detected by the LAMP assay within 60 min without cross-reaction to any other bacteria. Furthermore, a total of 135 nasal swab samples were tested directly using our LAMP assays, the previously reported LAMP assay, conventional PCR assay without pre-culture and comparing standard culture methods. The improved LAMP assay showed sensitivity and specificity of 97.2% and 90.9%, respectively (with a kappa coefficient of 0.8231), and the sensitivity of our revised LAMP assay was increased compared to existing methods.

The effect of depopulation and restocking on reproductive and growth performances on Japanese commercial swine farms

This study compared the reproductive and growth performances of pigs before and after depopulation and restocking after a foot-and-mouth disease outbreak in Japan. Data for the time period before and after depopulation and restocking were obtained from three farrow-to-finish farms. As a result of depopulation and restocking, hygiene levels were improved, and common infectious diseases, such as porcine reproductive and respiratory syndrome and Aujeszky’s disease, remained undetected on the farms. Compared with before depopulation, reproductive and growth performances improved after depopulation; the number of total pigs born was higher, the postweaning mortality rate was lower, and the age at slaughter was lower (P<0.05). In summary, depopulation and restocking improved the reproductive and growth performances of pigs.

Use of Direct LAMP Screening of Broiler Fecal Samples for Campylobacter jejuni and Campylobacter coli in the Positive Flock Identification Strategy

Rapid identification of Campylobacter-positive flocks before slaughter, following freezing and heat treatment for the Campylobacter-positive carcasses at the slaughterhouses is an effective control strategy against foodborne campylobacteriosis. We evaluated a loop-mediated isothermal amplification (LAMP) assay for the direct screening of naturally contaminated chicken cloacal swabs for C. jejuni/C. coli to compare this assay with conventional quantitative culture methods. In a comparison study of 165 broilers, the LAMP assay showed 82.8% (48/58 by conventional culture) sensitivity, 100% (107/107) specificity, 100% (48/48) positive predictive value (PPV), and 91.5% (107/117) negative predictive value (NPV). In a comparison of 55 flocks, LAMP showed 90.5% (19/21) sensitivity, 100% (34/34) specificity, 100% (19/19) PPV, and 94.4% (34/36) NPV. In the cumulative total of 28 farm-level comparisons, LAMP showed 100% (12/12) sensitivity, 100% (16/16) specificity, 100% (12/12) PPV, and 100% (16/16) NPV. The LAMP assay required less than 90 min from the arrival of the fecal samples to final results in the laboratory. This suggests that the LAMP assay will facilitate the identification of C. jejuni/C. coli-positive broiler flocks at the farm level or in slaughterhouses before slaughtering, which would make it an effective tool in preventing the spread of Campylobacter contamination.

An analysis of factors affecting production performance in broiler flocks on Japanese commercial farms

Abstract 1. The present study determined descriptive values of the main production measurements of flocks and assessed the relationship between these measurements and related management factors in Japanese commercial broiler farms. 2. The data set included 5060 flock records from 183 farms. The production index was calculated as follows: liveability × average daily gain/feed conversion ratio × 10. Management factors included in the analysis were broiler breeder age, the time interval between successive flocks, the season of placement and stocking density. 3. The mean (±SD) production index was 283.9 ± 28.83. Management factors significantly associated with a decreased production index were low broiler breeder age, flocks placed in summer and high stocking density (P < 0.05). 4. In regard to an interaction for the production index, flocks with high stocking density had a lower production index than those with low stocking density in flocks with a low broiler breeder age (P < 0.05). In summer, flocks with a short time interval between successive flocks had a lower production index than those with an intermediate or long time interval (P < 0.05). 5. The present study identified factors related to flock performance. The knowledge obtained from this analysis will contribute to improve flock performance by optimising management.

Effects of zinc supplementation on Shiga toxin 2e-producing Escherichia coli in vitro

Swine edema disease is caused by Shiga toxin (Stx) 2e–producing Escherichia coli (STEC). Addition of highly concentrated zinc formulations to feed has been used to treat and prevent the disease, but the mechanism of the beneficial effect is unknown. The purpose of the present study was to investigate the effects of highly concentrated zinc formulations on bacterial growth, hemolysin production, and an Stx2e release by STEC in vitro. STEC strain MVH269 isolated from a piglet with edema disease was cultured with zinc oxide (ZnO) or with zinc carbonate (ZnCO3), each at up to 3,000 ppm. There was no effect of zinc addition on bacterial growth. Nonetheless, the cytotoxic activity of Stx2e released into the supernatant was significantly attenuated in the zinc-supplemented media compared to that in the control, with the 50% cytotoxic dose values of 163.2 ± 12.7, 211.6 ± 33.1 and 659.9 ± 84.2 after 24 hr of growth in the presence of ZnO, ZnCO3, or no supplemental zinc, respectively. The hemolytic zones around colonies grown on sheep blood agar supplemented with zinc were significantly smaller than those of colonies grown on control agar. Similarly, hemoglobin absorbance after exposure to the supernatants of STEC cultures incubated in sheep blood broth supplemented with zinc was significantly lower than that resulting from exposure to the control supernatant. These in vitro findings indicated that zinc formulations directly impair the factors associated with the virulence of STEC, suggesting a mechanism by which zinc supplementation prevents swine edema disease.