Borrelia burgdorferi Sensu Lato in an Endemic Environment: Wild Sika Deer (Cervus nippon yesoensis) with Infected Ticks and Antibodies

Ticks and blood samples were collected from wild sika deer (Cervus nippon yesoensis) during a hunting season (August to October) of 1991 at a selected location in Hokkaido, Japan. Ixodes persulcatus (adult and nymph) and I. ovatus (adult) were the common ticks on sika deer. Spirochetes were detected in the midgut of the ticks by the indirect peroxidase-conjugated antibody staining method and by dark-field microscopy after cultivation. By the reactive pattern of monoclonal antibodies, isolates were considered to belong to Borrelia garinii or B. japonica. In an antibody test, the percentage of seropositive deer was 69.0%. Most of the adult sika deer were positive for antibodies to the spirochetes. There are significant age-dependency in antibody level and seropositive rate. The surveillance of deer should be valuable in monitoring the transmission risk of B. burgdorferi sensu lato in nature.     

Serological survey for Lyme disease in sika deer (Cervus nippon yesoensis) by enzyme-linked immunosorbent assay (ELISA).

Seventy-six wild sika deer (Cervus nippon yesoensis) from areas endemic for Borrelia burgdorferi during 1988 to 1989 had the IgG antibody to the bacteria in enzyme-linked immunosorbent assay. The percentage of seropositive deer was 25.0% to strain HO14 and 22.4% to strain HP3, respectively. Specific IgG antibody titers were high in summer but low in winter. In summer, seropositive deer were 75.0%. Similar results were obtained in serum specimens obtained monthly from 4 farmed deer. It was suggested that the sika deer could be one of the wild reservoirs for B. burgdorferi in Hokkaido, Japan. The transmission risk of B. burgdorferi is considered to be greatest during late spring to early summer.     

Resistance of Pseudomonas pseudomallei to Normal Human Serum Bactericidal Action

The effect of human normal serum (HNS) on Pseudomonas pseudomallei was determined. It is apparent from our data that the organism is resistant to the normal serum bactericidal mechanism. Ancillary experiments to confirm this serum-resistant property of P. pseudomallei were done by examining the effects of growth phase conditions of the bacteria (i.e., logarithmic and stationary phases) and different buffered systems used as diluent in our bactericidal assay. Results obtained showed similar degree of resistance to serum bactericidal killing by 5 strains of the organisms tested. The possible survival advantage of serum-resistant property to P. pseudomallei as bacterial pathogens known to invade the blood stream is discussed.     

Prevalence of Antibodies to Borrelia Spirochetes among Wild Small Rodents in Central and Western Japan

Field rodents serve as a reservoir for Lyme disease spirochetes. To evaluate the antibody responses of rodents against different Borrelia species in relation to fauna of vector ticks feeding on them, we examined 272 sera of wild rodents, Apodemus speciosus, A. argenteus, and Eothenomys smithii, obtained in 27 locations in central and western Japan from 1981 to 1994. As to prevalences by rodent species using immunoperoxidase test, A. speciosus, A. argenteus and E. smithii showed 29.4%, 11.6% and 30.8% reactivity to Borrelia japonica, 10.7%, 7.2% and 3.8% to B. afzelii, 0.6%, 1.4% and 0% to B. garinii, and 14.7%, 7.2% and 11.5% to an unknown Borrelia species designated as It type, respectively. Each antibody to B. japonica, B. afzelii and B. sp. It type was detected widely both in central and western Japan, but the antibody to B. garinii was scarcely detectable in any area and rodent species examined. Apodemus mice in high mountain altitudes tend to have antibody to B. afzelii or B. japonica, and those in lower altitudes tend to have B. japonica or B. sp. It type. All 13 Apodemus mice from which B. japonica or B. sp. It type were isolated showed higher titers of antibodies to each homologous Borrelia species. The present results indicate that these antibody prevalences among rodents may be associated with dominant Ixodes ovatus and sporadic I. persulcatus on the mainland of Japan, and that Apodemus mice may not be an efficient reservoir for B. garinii. Such a serosurvey is a useful measure to evaluate the natural distribution of the pathogen.     

Glycosylated hemoglobin as a stable alternative to serum glucose in white-tailed deer

We compared serum glucose concentration and percent glycosylated hemoglobin (GH) in captive and wild white-tailed deer (Odocoileus virginianus) to determine stability of glucose relative to GH. Temporal changes in levels of serum glucose and GH were ascertained from serial blood samples collected from three captive deer over a 2-week period. State of glycemia also was determined for 17 wild deer that were collected from three populations in southeastern Oklahoma and southwestern Arkansas (USA). Concentration of serum glucose of captive deer decreased (P = 0.04) from 190.4 to 155.8 mg/dl over the 2 weeks; percent GH did not differ temporally (P = 0.30). Percent GH of wild deer did not differ (P = 0.23) when deer were separated into 2 groups (high and low state of glycemia) based on the median serum glucose concentration. We found a significant difference (P = 0.04) in percent GH among wild deer populations; serum glucose concentration did not differ (P = 0.72) among populations. Our results indicate that percent GH is more stable than serum glucose concentration and may be useful in population comparisons of nutritional condition.     

The Stability of Complement-Mediated Bactericidal Activity in Human Serum against Salmonella

The complement cascade includes heat-labile proteins and care is required when handling serum in order to preserve its functional integrity. We have previously used a whole human serum bactericidal assay to show that antibody and an intact complement system are required in blood for killing of invasive isolates of Salmonella. The aim of the present study was to evaluate the conditions under which human serum can be stored and manipulated while maintaining complement integrity. Serum bactericidal activity against Salmonella was maintained for a minimum of 35 days when stored at 4°C, eight days at 22°C and 54 hours at 37°C. Up to three freeze-thaw cycles had no effect on the persistence of bactericidal activity and hemolytic complement assays confirmed no effect on complement function. Delay in the separation of serum for up to four days from clotted blood stored at 22°C did not affect bactericidal activity. Dilution of serum resulted in an increased rate of loss of bactericidal activity and so serum should be stored undiluted. These findings indicate that the current guidelines concerning manipulation and storage of human serum to preserve complement integrity and function leave a large margin for safety with regards to bactericidal activity against Salmonella. The study provides a scheme for determining the requirements for serum handling in relation to functional activity of complement in other systems.     

Comparison of OspA Serotypes for Borrelia burgdorferi Sensu Lato from Japan,Europe and North America

Sixty-one Borrelia burgdorferi sensu lato strains from various sources (ticks, human, and wild animals) in Japan and two strains from ticks in Far Eastern Russia were classified on the basis of reactivity with 16 monoclonal antibodies (mAb) to outer surface protein A (OspA) and by DNA-DNA hybridization assay. Eleven OspA serotypes (J1 to J11) were recognized among the Japanese and the Far East Russian isolates (serotypes J1 to J9 were identified as B. garinii, serotype J10 was identified as B. afzelii, and serotype J11 corresponded to B. japonica), whereas 7 OspA serotypes for North American and European isolates previously reported (Bettina Wilske et al, J. Clin. Microbiol. 31:340-350, 1993) were not observed except for OspA serotype 2 which showed identical reactivity with OspA serotype J10. This finding provides helpful information for understanding the geographical distribution of Lyme disease borrelia and the development of vaccine and diagnostic tests. In conclusion: 1. B. burgdorferi sensu stricto has not been observed in Japan, 2. Japanese B. afzelii isolates are closely related to those from Europe, 3. B. garinii isolates from Japan are highly heterogeneous and apparently different from European B. garinii isolates.     

同域分布的野猪与狍活动节律及与月光周期的关系

完全的竞争者不能共存,物种间生态位分化是同域物种长期稳定共存的基础。不同物种在同一分布区时间生态位的分化对其共存至关重要。为研究同域分布物种的共存机制,在2018年11月到2021年7月利用远红外相机监测技术对穆棱东北红豆杉国家级自然保护区同域分布的野猪(Sus scrofa)和狍(Capreolus pygargus)进行了野外监测研究,并利用核密度估计方法和雅各布斯选择指数(JSI)对日活动节律和一段时间内的活动周期选择进行了评估,同时还利用非参数检验评估了2个物种的活动与月光周期的关系。研究结果表明,狍不管在全年还是在不同的季节,日活动节律均为双峰模式,而野猪的活动模式表现出明显的可塑性,在全年和冷季为单峰型,暖季为双峰型,二者在日活动节律上的重叠程度较高(Dhat>0.59),在暖季的重叠系数最高(Dhat=0.65)。此外,在时间段的选择上,狍在全年和冷季更喜欢在黎明和黄昏活动(0.170.32),而野猪活动在全年和冷季对白天和黄昏的选择更多(JSI>0.3),暖季则更喜欢在白天活动(J...     

Genetic Diversity of Borrelia burgdorferi Sensu Lato Isolated in Far Eastern Russia

One-hundred and fifty-seven Borrelia isolated from adult ticks, Ixodes persulcatus, and wild rodents, Clethrionomys rufocanus and Apodemus peninsulae, in the far eastern part of Russia were characterized and identified by restriction fragment length polymorphism (RFLP) of the 5S-23S rRNA intergenic spacer. Some isolates showed unique RFLP patterns and were determined as Borrelia garinii on the basis of a sequence analysis of the intergenic spacer amplicon and reactivity with species-specific monoclonal antibodies (MAbs). 86.5 and 12.7% of the tick isolates, and 74.2 and 12.9% of the rodent isolates were determined as Borrelia garinii and Borrelia afzelii, respectively, but no Borrelia burgdorferi sensu stricto was detected. This finding is similar to the results obtained from Borrelia surveys of I. persulcatus and wild rodents in Hokkaido, Japan.     

An in vitro synergetic evaluation of the use of nisin and sodium fluoride or chlorhexidine against Streptococcus mutans

The objective of this study is to investigate the synergetic action between nisin and sodium fluoride or chlorhexidine against Streptococcus mutans, a primary cariogenic pathogen. In the antibacterial assay, a synergetic effect on S. mutans was found between nisin and sodium fluoride, but there was no interaction between nisin and chlorhexidine by the checkerboard, the fractional inhibitory concentration (FIC) and the fractional bactericidal concentration (FBC) tests. S. mutans survival rates showed a significant decline after treatment with a combination of nisin and sodium fluoride in a time-kill study. Scanning electron microscopy showed that the damage to S. mutans with the combined nisin and sodium fluoride treatment was the most severe among all of the different single and combined antimicrobial treatments. Furthermore, in the antibiofilm test, nisin in combination with sodium fluoride produced a stronger bactericidal effect on a S. mutans biofilm for 4 h and 16 h compared with sodium fluoride alone by confocal laser scanning microscopy. Nisin in combination with sodium fluoride exerted a high bactericidal effect on S. mutans and thereby has the potential to be used as an effective drug combination to prevent dental caries.     

甘肃兴隆山保护区野生马麝分布、数量特征及影响因素

综合采用样线法、粪堆计数法及重复调查法对甘肃省兴隆山国家级自然保护区的野生马麝（Moschus chrysogaster）进行了种群调查,结合生境分析,确定了其种群分布、数量特征及影响因素。结果表明,兴隆山保护区分布有野生马麝（1159±275）头,平均种群密度为（3.51±0.83）头/km²;各植被类型生境中的野生麝种群数量及密度存在差异,灌丛生境分布有70%的野生马麝种群,达（807±170）头,种群密度为（6.49±1.63）头/km²;针叶林种群密度最大,达（8.85±83.25）头/km²,分布有野生麝（123±45）头;针阔混交林分布最少,仅（41±15）头,种群密度为（5.00±1.84）头/km²;人工林生境无野生马麝分布。保护区各植被类型生境中的野生马麝种群分布差异反映了马麝对适宜生境功能的需求,食物、保温和隐蔽性是制约野生马麝冬季分布和种群数量的关键因素。此外,因生境及人为干扰强度的不同,保护区各区域的野生马麝种群分布存在差异。建议通过减少人为干扰、地表植被管理及人工林管理优化等措施增加生境适宜性,促进兴隆山自然保护区野生马麝种群的快速恢复和增长。     

Population dynamics in a guild of four Mediterranean ungulates: density‐dependence,environmental effects and inter‐specific interactions

Population fluctuations in ungulates are driven by both intrinsic and extrinsic factors. Available information, however, mainly refers to arctic, temperate and African ungulate populations, while the dynamics of Mediterranean species, exposed to a milder climate, is known to a much lesser extent. Here we studied the population dynamics of four wild ungulate species in the Castelporziano Preserve near Rome, Italy, as obtained from detailed bag counts from hunting drives during the period 1878–1986: the Italian roe deer Capreolus capreolus italicus, the Maremma wild boar Sus scrofa majori (both endemic to Italy), the native red deer Cervus elaphus, and the alien fallow deer Dama dama. We also considered the effects of the presence of another alien ungulate, the nilgai Boselaphus tragocamelus. This ungulate community experienced an accidental ‘removal experiment’ when, during World War II, red deer and nilgai were exterminated. This event and the length of the time series allowed us to test two main hypotheses: 1) that the complexity level of the ungulate community affects the strength of intra‐ and inter‐specific competition; and 2) that in Mediterranean environments intra‐ and inter‐specific interactions are stronger than climate forcing. Statistical methods ranged from state‐space‐modelling, GLM analysis and structural equation models. The results indicated that direct intra‐specific density dependence played a relevant role for all species, and was stronger after the removal. A complex pattern of species interactions was however revealed; fallow deer had a negative effect on roe deer population, while roe deer had an apparent positive effect on red deer and wild boar, possibly mediated by environmental factors. Nilgai appeared to facilitate all deer species. The results of the analysis also confirmed that at present climate appears to play a minor role with respect to density dependence; however, the increasing aridity of the Mediterranean area could change this picture in coming decades.     

Correlation of immune assays with protection in rainbow trout, Salmo gairdneri, immersed in Vibrio bacterins

One hundred kg of Rainbow trout, Salmo gairdneri, weighing 80–100g each were immersed in a formalized, bivalent vaccine of Vibrio anguiliárum (VA) and Vibrio ordalii (VO), at a1/10 dilution for 30s at 10°C. The protection levels achieved at 8 and 10 weeks post vaccination were assessed by intraperitoneal challenge with 10⁵ live V. anguillarnm or V. ordalii. No decline in protection due to antigen depletion was found between batches throughout the procedure. The protection conferred by one component of the vaccine was slightly better than the other (80% against VO and 42 % against VA). A number of measures of immunity were monitored before and after challenge. These included serum and mucus antibody litres, bactericidal effects of serum and mucus and phagocytosis by peritoneal exudate cells. Serum antibody was present (peak litre of-log₂ 3.25 for VO, and 4.4 for VA) but had declined to background levels by the time of challenge, whereas bactericidal effects and phagocytosis rate of 40% were not increased by vaccination. The most likely immune mechanism responsible for the observed protection was discussed and suggested to be a cellular mechanism where serum antibody acts as an opsonin to increase phagocytosis.     

Seroprevalence of Toxoplasma gondii in wild boars (Sus scrofa leucomystax) and wild sika deer (Cervus nippon) in Gunma Prefecture, Japan

The ingestion of undercooked meat from wild animals can be a source of Toxoplasma gondii infection in humans and other animals. In this study, we determined the seroprevalence of T. gondii infection in 175 wild boars (Sus scrofa leucomystax) and 107 wild sika deer (Cervus nippon) hunted in 2004–2007 in Gunma Prefecture, Japan, by using a commercial latex agglutination test (LAT). Antibodies (LAT, 1:64 or higher) to T. gondii were found in 6.3% of wild boars and 1.9% of sika deer. This is the first record of T. gondii infection in wild deer in Japan, and deer and wild boar meat should be cooked well before human consumption.     

张广才岭南部东北虎猎物种群冬季营养容纳量研究

猎物匮乏是影响东北虎（Panthera tigris altaica）种群恢复的关键因素之一。容纳量研究是开展东北虎猎物恢复工作的必要前提。通过Maxent模型、聚类分析和训练随机树分类等方法,结合调查数据,预测了吉林省张广才岭南部黄泥河林业局东北虎主要猎物的适宜栖息地空间分布,解译了植被类型,在此基础上基于不同植被类型动物可采食部分代谢能、不同生境等级食物可利用率、马鹿（Cervus elaphus）和狍（Capreolus capreolus）生境等级重叠情况以及动物冬季能量需求,分析了东北虎猎物的冬季营养容纳量。结果表明：黄泥河林业局狍、野猪（Sus scrofa）和马鹿的适宜栖息地分别占研究区域总面积的52.8%、40.7%和25.4%;从猎物获取能量来看,以山杨（Populus davidiana）、桦树（Betula）、核桃楸（Juglans mandshurica）为主的植被类型是马鹿、狍可获得能量较多的生境,以蒙古栎（Quercus mongolica）、核桃楸为主的植被类型是野猪可获得能量较多的生境。东北虎猎物种群的综合冬季营养容纳量为574只马鹿（0.29只/km²）,7016只狍（3.54只/km²）,4785只野猪（2.38只/km²）。     

Identification of Borrelia burgdorferi Isolated in Korea Using Outer Surface Protein A (OspA) Serotyping System

Two characteristic strains (935T, 934U) of B. burgdorferi isolated from Ixodes persulcatus and a wild rodent (Apodemus agrarius) in Korea were selected and analyzed by an immunoblot method using the monoclonal antibodies directed to different epitopes of outer surface protein A (OspA). The reactive pattern of strain 934U with these monoclonal antibodies was identical to that of strains belonging to B. afzelii and that of strain 935T was different from other isolates. Monoclonal antibody (5TEE3) which is specific to strain 935T did not react with any other Western and Japanese isolates. So, it was suggested that there exist at least two groups of B. burgdorferi in Korea. One could be classified as B. afzelii and the other is a divergent group from three known species of B. burgdorferi sensu stricto, B. garinii and B. afzelii.     

Ecological characterization of murine gammaherpesvirus 68 and it’s epidemiological implications

Murine gammaherpesvirus 68 (MHV-68) is a natural pathogen of free-living murid rodents, isolated from a bank vole (Clethrionomys glareolus) in Slovakia. It belongs to the family Herpesviridae, the genus Rhadinovirus, the species Murid herpesvirus 4. Neutralizing serum MHV-68 antibodies were found in animals of other species living with infected rodents in the same biotope (fallow deer, wild boars, deer, sheep, foxes, and mouflons) as well as in humans (laboratory personnel working with MHV-68 or wild rodents, hunters, general population). In this study, the shedding of MHV-68 in the nature was investigated on the model of laboratory mice experimentally infected with the virus. The virus was first detected in breast milk and lactic glands at day 4, in urine and salivary glands at day 9, and in saliva and tear glands at day 14 p.i. Out of intranasal, peroral, intramuscular, intraperitoneal, and subcutaneous routes of infection the first two proved most effective.     

Wolf predation and snow cover as mortality factors in the ungulate community of the Bialowieża National Park,Poland

Summary Wolf-ungulate interactions were studied in the pristine deciduous and mixed forests of the Bialowiea National Park in 1985–1989. The study period included two severe and two mild winters. The community of ungulates inhabiting Bialowiea National Park consisted of red deer Cervus elaphus, 55% of all ungulates; wild boar Sus scrofa, 42%; and roe deer Capreolus capreolus, moose Alces alces, and European bison Bison bonasus, about 1% each. The average size of red deer groups increased from 2.7 (SD 2.35) in spring and summer to 6.9 (SD 6.84) in autumn and winter. In winter the group size of red deer was positively correlated with the depth of snow cover and negatively correlated with the mean daily temperature. Average group size of wild boar did not change significantly between seasons; it was 6.8 (SD 5.16) in spring and summer and 5.7 (SD 4.67) in autumn and winter. Analysis of 144 wolf scats showed that wolves preyed selectively on red deer. In October–April, Cervidae (mostly red deer) constituted 91% of biomass consumed by wolves, while wild boar made up only 8%. In May–September deer formed 77% of prey biomass, and the share of wild boar increased to 22%. In all seasons of the year wolves selected juveniles from deer and boar populations: 61% of red deer and 94% of wild boar of determined age recovered from wolves' scats were young <1 year old. Analysis of 117 carcasses of ungulates found in Bialowiea National Park showed that predation was the predominant mortality factor for red deer (40 killed, 10 dead from causes other than predation) and roe deer (4 killed, none dead). Wild boar suffered most from severe winter conditions (8 killed, 56 dead). The percentage of ungulates that had died from undernutrition and starvation in the total mortality was proportional to the severity of winter.     

新疆塔里木马鹿粪便激素含量季节性变化的初步研究

目的:检测雄性塔里木马鹿的睾酮,雌鹿的雌二醇、雌三醇、孕酮等粪便激素含量,并比较不同月份之间的变化。方法:利用放射免疫分析法(RIA)在且末和尉犁县马鹿场采集塔里木马鹿粪便,检测了2004年2～12月间塔里木马鹿粪便中的激素水平。结果:粪便性激素水平有明显的季节变化(P<0.05)。雄鹿粪便睾酮水平月间差异显著(P<0.05),浓度在8月有2个高峰。雌鹿粪便雌二醇在3月和6月有2个高峰;粪便孕酮水平在3月有2个高峰,在8月有1个高峰;雌三醇差异不显著,但4月的最高值也显著高于其他各时间点(P<0.05)。结论:利用动物粪便研究激素具有对保护动物完全无伤害、材料收集容易等优点,并且有助于评估动物青春期的健康状况、判断动物排卵方式、早孕检查、产前预测等,因而是目前研究野生珍稀动物性激素的一种较为理想的方法。     

Survey of Pestivirus infection in wild and domestic ungulates from south-western Italian Alps

The transmission of pestiviruses between domestic and wild ruminants has not been documented in communal alpine pastures shared between wildlife and livestock. The aim of this study was to investigate the role of domestic and wild ungulates species from Varaita Valley (SW Italian Alps) in the epidemiology of Pestivirus infections. Sera from free-ranging alpine chamois (Rupicapra rupicapra) and roe deer (Capreolus capreolus) were collected from 1994 to 2009 and 2001 to 2009, respectively. Also, sera from cattle and sheep sampled in 2009 were studied. Sera were tested for the presence of antibodies against pestivirus with an ELISA assay. Sera from positive animals were subsequently tested with a comparative virus neutralisation test using the BVDV-NADL and BDV-137/4 strains. Sera were tested for the presence of pestiviral antigen and the presence of viral RNA with a commercial ELISA assay and RT-PCR. Antibodies against Pestivirus were detected in 132 out of 312 (42%) chamois, in 30 out of 175 (17%) cattle and 6 out of 24 (25%) sheep. No antibodies were found in roe deer. No Pestivirus antigen or RNA was detected in any of the samples. Results indicate circulation of pestiviruses among the studied chamois, cattle and sheep populations. However the role of wild ungulates in the dynamics of Pestivirus infection is still unknown and monitoring the presence of these viruses in wild ungulates would be of importance, especially in the chamois population, where pestiviruses seem to circulate extensively.