Glyphosate biodegradation and potential soil bioremediation by Bacillus subtilis strain Bs-15.

Glyphosate and glyphosate-containing herbicides have an adverse effect on mammals, humans, and soil microbial ecosystems. Therefore, it is important to develop methods for enhancing glyphosate degradation in soil through bioremediation. We investigated the potential of glyphosate degradation and bioremediation in soil by Bacillus subtilis Bs-15. Bs-15 grew well at high concentrations of glyphosate; the maximum concentration tolerated by Bs-15 reached 40,000 mg/L. The optimal conditions for bacterial growth and glyphosate degradation were less than 10,000 mg/L glyphosate, with a temperature of 35°C and a pH of 8.0. Optimal fermentation occurred at 180 rpm for 60 h with an inoculum ratio of 4%. Bs-15 degraded 17.65% (12 h) to 66.97% (96 h) of glyphosate in sterile soil and 19.01% (12 h) to 71.57% (96 h) in unsterilized soil. Using a BIOLOG ECO plate test, we observed no significant difference in average well color development values between the soil inoculated with Bs-15 and the control soil before 72 h, although there was a significant difference (P < 0.01) after 72 h. In the presence of Bs-15, the 5 functional diversity indices (Shannon index, Shannon uniformity, Simpson index, McIntosh index, and McIntosh uniformity) were greater (P < 0.01) compared with the control soil. These results indicate that Bs-15 could be used to alleviate contamination from glyphosate-containing herbicides, increasing the microbial functional diversity in glyphosate-contaminated soils and thus enhancing the bioremediation of glyphosate-contaminated soils.

Preparation of monoclonal antibody of anti-feline calicivirus and establishment of double-antibody sandwich enzyme-linked immunosorbent assay detecting method.

This study aimed to prepare monoclonal antibody of feline calicivirus (FCV) and identify its basic biological characteristics. Saturated ammonium sulfate precipitation, combined differential centrifugation, and cesium chloride density gradient centrifugation were used for the purification of FCV. The purified FCV was used as antigen to immunize BALB/c mice. The hybridoma lines of anti-FCV monoclonal antibodies were established using cell fusion and hybridoma screening techniques. The subtypes of the monoclonal antibody were identified. The results showed that 3 strains of hybridoma cell lines stably secreted anti-FCV monoclonal antibody; they were named as D8, E5, and H4. The D8 and E5 were IgM subtype antibodies, and H4 was IgG2b subtype antibody. The monoclonal antibody obtained shared no cross-reactivity with feline parvovirus, canine parvovirus, and canine distemper virus. According to the different recognition sites of 2 monoclonal antibodies E5 and H4 to the FCV, they were used to coat microtiter plates and prepare 2 enzyme-labeled secondary antibodies to establish double-antibody sandwich enzyme-linked immunosorbent assay detecting method.

Epidemiological and aetiological evidence for transmission of Lyme disease by adult Ixodes persulcatus in an endemic area in China.

Involvement of adult Ixodes persulcatus ticks in the transmission of Lyme disease in Hailin County, Heilongjiang Province, China, is reported. In 1986 from April through August adult I. persulcatus was the dominant tick in this endemic area with an infection rate of 43% for the Lyme disease spirochaete, Borrelia burgdorferi. The incidence of Lyme disease cases presenting the symptom of erythema chronicum migrans (ECM) within this area was correlated with the seasonal abundance of adult I. persulcatus and the number of people bitten by ticks. The frequency of ECM formation in all age groups varied and was associated with the frequency of tick bites. In addition, a strain of B. burgdorferi was isolated from a pool of six female I. persulcatus collected from this area. We demonstrate that the seasonal abundance of adult I. persulcatus and its frequent attachment to humans result in the spring and summer transmission of Lyme disease in this endemic area. The role of immature I. persulcatus in Lyme disease transmission is apparently minimal.

Clinical manifestations and epidemiological characteristics of Lyme disease in Hailin county, Heilongjiang Province, China.

Clinical manifestations and epidemiological characteristics of Lyme disease in Hailin county, Heilongjiang Province, China have been reported. The clinical picture of erythema chronicum migrans (ECM) is variable. ECM in the form of annular erythematous patch is uncommon. It is an extensive and indurated lesion. In some instances, a vesicle or necrosis appears in the center of the lesion. Secondary erythema may present in some patients. The neurologic abnormalities consist of meningitis, facial palsy, and polyneuritis. Cardiac abnormalities are rare. In addition, there were cases with lymphadenosis benigna cutis (LABC), which had heretofore only been reported in Europe. The attack rate of ECM is 8.4%. There was a significant sex difference, and most cases occurred in May and June. All patients had a history of tick bite. The prevalence rates of neurologic abnormalities and arthritis were 4.6% and 6.6%, respectively. Three strains of spirochete that are closely related to Borrelia burgdorferi were isolated from Ixodes persulcatus ticks and facial palsy patients. From the above results it is concluded that a focus of Lyme disease exists in this region.

Borrelia burgdorferi infection may be the cause of sarcoidosis.

Serum antibody to Borrelia burgdorferi was measured in 33 patients with sarcoidosis which was confirmed clinically and pathologically. The results showed that 81.8% of the patients were positive for anti-B. burgdorferi antibody. In addition, a strain of B. burgdorferi was isolated from a patient's blood. Fifteen patients received ceftriaxone 2g per day or penicillin 12 million U per day. The antibody titers of the patients decreased to nearly normal levels rapidly. Serum angiotensin converting enzyme (SACE) turned to normal range after the treatment. According to the findings mentioned above, we consider that B. burgdorferi infection may be the cause of sarcoidosis and sarcoidosis might be a specific type of Lyme disease.

[Facial palsy in Lyme disease].

This paper reports twenty four cases of facial palsy in Lyme Disease for the first time in the forest region. Most cases had history of tick bite. Two strains of spirochete were isolated from the blood of 7 patients with facial palsy. These spirochetes were immunologically closely related to B. burgdorferi. Eighteen serum samples of facial palsy were tested by indirect fluorescence assay, using B. burgdorferi B-31 as antigen. In serum, IgG antibody titers of 1:64 to 1:2048 were found in 50% of the cases. We suggest that neurologists should consider the possibility of Lyme Disease in the diagnosis of facial palsy.

Sero-epidemiology of Lyme disease in an endemic area in China.

A sero-epidemiological investigation on Lyme disease was carried out in a forestry center of Hailin County, Heilongjiang Province, China. A total of 381 participants including forestry workers and their dependents completed questionnaires and had blood samples taken for detection of antibody against Borrelia burgdorferi by indirect immunofluorescent assay. Of 381 participants, 250 (65.6%) had a history of tick bites between May and July, 1987, and 379 (99.5%) at some time in the past, 56 (14.7%) developed erythema migrans at the site of tick attachment, 138 (36.2%) had late manifestations of Lyme disease, and 101 (26.2%) were seropositive. There was a significant difference in the seropositive rate between the persons with tick bites and those without tick bites in 1987 (P < 0.01). The titers in the older age group (over 30) were slightly higher than those of the younger age group (under 30). No relationship between the prevalence of Lyme manifestations by tick bites and the result of serological test was observed. Further investigations are necessary to assess the actual prevalence and incidence of infections using antigen of Lyme disease borreliae isolated in China.

Host associations and seasonal abundance of immature Ixodes persulcatus (Acari: Ixodidae) in a Lyme-disease-endemic area in China.

Ticks of nine small mammal species were studied in an area of endemic Lyme disease in Hailin County, Heilongjiang Province, from 1 May to 10 October 1986. The bulk (93%) total small mammal captures were Apodemus speciosus (45%), Clethrionomys rufocanus (31%), A. agrarius (11%) and Eutamias sibiricus; (5%): these host species accounted for 97% of the 162 ticks collected. Ixodes persulcatus Schulze were most abundant on E. sibiricus (0.64 larvae and 0.08 nymphs/squirrel), and less abundant on A. agrarius (1.16 larvae/mouse), A. speciosus (0.38 larvae/mouse) and C. rufocanus (0.37 larvae and nymphs/vole). The seasonal abundance pattern of larval I. persulcatus on four small mammal species appears as a bimodal peak (June and September), but nymphal I. persulcatus has a monomodal peak during spring. The prevalence of immature I. persulcatus on four small mammal species may play an important role in the epidemiology of Lyme disease in Hailin County.