Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus)

Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.     

Developing and testing a habitat suitability index model for Korean water deer (Hydropotes inermis argyropus) and its potential for landscape management decisions in Korea

Geographic information system (GIS) and landscape-level data offer a new opportunity for modeling and evaluating the quality of wildlife habitats. Models of habitat quality have not been developed for some species, and existing models could be improved by incorporating updated information on wildlife–habitat relationships and habitat variables. We developed a GIS-based habitat suitability index (HSI) model for the Korean water deer (Hydropotes inermis argyropus), which often causes human–wildlife conflicts in the Chungnam Province of Korea because of industrialization and urbanization. The model is based on logistic regression analysis, which addresses the impact of multiple habitat variables, such as habitat components, topographic characteristics, and human disturbances. The model yielded a p-value of .289 (χ²?=?9.672) and 65.4% correct prediction level with the overall observation–prediction comparison data. The model demonstrated that a large portion of the province (61.6%) could be regarded as a poor habitat (mean HSI value of the province?=?0.22), while the current habitats of the province could be considered of moderate quality (mean HSI value?=?0.31). In addition, the chance of observation of the deer increases as the HSI level increases, which means that the model yields a good predictive power. Lastly, we used the model to produce a habitat suitability map. Our HSI model enabled us to quantify habitat preferences, which could be the basis for decision-making on habitat protection, mitigation, and enhancement of the Korean water deer. The proposed model is also applicable for improving and enhancing the existing management practices, as well as for establishing an effective wildlife protection policy.     

舟山群岛獐的食性研究

利用野外观察、粪便显微组织学分析和投喂实验等方法对舟山群岛獐的食性进行了研究。结果表明,研究地的獐所食植物共有137种,隶属于6l科l15属,其中豆科植物在取食种类和取食频次上均多于其它科植物。尽管獐对植物科和种的选择在四季间没有显著差别,但取食科和种的数量依然从春天到冬天呈递减趋势,并且冬天与其它3个季节差别较大。獐的食物中,每个季节都含有非禾草类草本植物、禾草类草本植物、木本植物和蕨类植物,且其食物组成在季节间亦无显著差异;除木本植物外,獐对其它3类植物的选择在季节间有显著差异。獐在冬天较多选择木本植物,而其它季节较多选择草本植物,但木本植物和非禾草类草本植物在四季食物组成中均占较高比例,而禾草类植物所占的比例很小。因此,笔者认为舟山群岛的獐应该偏向于嫩食者。     

獐在吉林省的重新发现

獐(Hydropotes inermis)在历史上曾广泛分布于中国的辽东半岛、华北平原及长江两岸,在1949年以后的多篇文献中没有发现对吉林省獐分布的记载。2017年12月至2018年3月,利用红外相机于吉林省白山市(原浑江市)白山原麝国家级自然保护区拍摄到獐有效照片21张,有效视频时长总计600 s。在后续的野外调查中,研究者现场拍摄到獐幼崽活体及大量獐的活动痕迹,确认獐在吉林省重新发现。     

Distribution, density, and habitat use of the Korean water deer (Hydropotes inermis argyropus) in Korea

Water deer (Hydropotes inermis) belong to the genus Hydropotes, which is ecologically well adapted for environments ranging from desert to forest. Water deer tend to occupy the richest areas between forest and grasslands. There are two distinct subspecies in far East Asia: one in China (H. inermis inermis), and the other in Korea (H. inermis argyropus). Despite being listed as vulnerable on the International Union for Conservation of Nature (IUCN) Red List, little is known about the water deer. The species occurs in most areas of the Korean peninsula, except for Seoul and Jeju Island. Kyunggi Province near Seoul showed the lowest rate (56%) due to urbanization. There was a high difference in their presence between inland (81%) and coastal (60%) areas . In addition, large cities (67%) showed much lower rates than did rural areas (83%) where human population size is relatively low. Water deer are distributed differently based on habitat type, with differences in mean density observed among lowland (6.93 ind./km²), mountainous (1.91), and urban (1.27) areas. There was also a difference in mean density between low-elevation and high-elevation areas. Finally, whereas the deer preferred landscapes with 20°–25° of slope and broadleaf forest (P < 0.01), they did not avoid areas with other types of slopes and habitats.     

獐(Hydropotes inermis)的警戒行为模式及逃跑起始距离的适应性变化

为了掌握獐的警戒行为特征并为重引入项目提供管理依据,以人为干扰源观察獐的警戒反应,发现其警戒模式包括听 (listen) 或扫视 (scan) ,盯视 (stare) ,走开 (walk away) ,跑开 (run away),吼叫 (bark) 和压脖 (stretch) 。利用逃跑起始距离对上海松江圈养 (自由采食) 獐、上海华夏圈养(人工饲喂) 獐和江苏盐城野生獐进行警戒性比较,得出：人工饲喂獐警戒性最小,野生獐警戒性最大。可通过野化训练提高人工饲喂獐的警戒性;人类活动、种群密度、围栏和区域面积可能影响獐的警戒性。     

Genetic Diversity of Chinese Water Deer (Hydropotes inermis inermis): Implications for Conservation

The Chinese water deer (Hydropotes inermis inermis) is endemic to China. Historically, the species was widely distributed, but now, habitat loss and poaching have reduced its range and number drastically. In order to provide useful information for its conservation, we have investigated the genetic diversity and population structure of the Chinese water deer by analyzing the 403 bp fragment of the mitochondrial DNA (mtDNA) control region (D-loop). Eighteen different haplotypes were detected in 40 samples. Overall, Chinese water deer have a relatively high-genetic diversity compared to other rare cervid species, with a haplotype diversity of 0.923±0.025 and nucleotide diversity of 1.318 ± 0.146%. No obvious phylogenetic structure among haplotypes was found for samples of different origin. An analysis of molecular variance (AMOVA) showed significant differentiation between the Zhoushan and the mainland population (F_ST= 0.088, P < 0.001; Φ _ST = 0.075, P = 0.043), which suggests that exchanges of individuals between Zhoushan and the mainland should be avoided. We also recommend that a breeding center be set up for the mainland population.     

Isolation and characterization of 12 microsatellite loci from Korean water deer (Hydropotes inermis argyropus) for population structure analysis in South Korea

The main goals of this study were to isolate microsatellites markers of Korean water deer (Hydropotes inermis argyropus) and understand the genetic status of the species in South Korea. Twelve new microsatellite loci were isolated and characterized to establish basic population genetic parameters for 45 H. i. argyropus specimens in South Korea. There were no significant regional or genetic structure differences between the mid-eastern and southwestern populations in South Korea according to the population genetic analyses. The number of alleles per locus ranged from two to 13 with an average of 6.08. Mean expected (H _E) and observed heterozygosity (H _O) were 0.622 and 0.533, respectively. Microsatellite variability was also not significant between the two regions (F _ST=0.012). These new markers should facilitate the future population genetics studies of Korean water deer and other closely related species.     

江苏盐城保护区獐的栖息地选择

1999年3月至2000年9月期间在江苏盐城保护区对獐的栖息地选择进行了研究。共记录了123个栖息地利用样方和300个任意样方,在样方中测定了5种生态因子(植被类型、植被高度、植被覆盖度、距水源距离和人为干扰距离)。不同季节獐对植被类型、植被高度和距水源距离的利用有显著差异,对植被覆盖度和人为干扰距离的利用有极显著差异。春季獐偏好选择白茅草滩、大穗结缕草滩、芦苇地和獐毛草滩;夏季主要选择碱蓬地、芦苇地和盐蒿草甸;秋季倾向于选择獐毛草滩、盐蒿草甸、藨草滩和大米草滩。在春、夏、秋季,獐都倾向于选择高度在90-110cm的植被。在不同季节獐都偏好选择植被覆盖度>90%的栖息地,主要选择距离水源200-599m之间的栖息地。     

Complete mitochondrial genome sequence of the Korean water deer Hydropotes inermis argyropus (Cervidae, Capreolinae)

The complete mitochondrial genome sequence of Hydropotes inermis argyropus consists of 13 protein-coding, 22 tRNA, and two rRNA genes, and 1 control region (CR). Three overlaps among the 13 protein-coding genes were found: ATP8/ATP6, ND4L/ND4, and ND5/ND6. The CR was located between the tRNA-Pro and tRNA-Phe genes and is 928 bp in length. The typical conserved domains, such as TAS and CSB, were identified in the CR.     

Intestinal Nematodes from Small Mammals Captured near the Demilitarized Zone,Gyeonggi Province,Republic of Korea

A total of 1,708 small mammals (1,617 rodents and 91 soricomorphs), including Apodemus agrarius (n = 1,400), Microtus fortis (167), Crocidura lasiura (91), Mus musculus (32), Myodes (= Eothenomys) regulus (9), Micromys minutus (6), and Tscherskia (= Cricetulus) triton (3), were live-trapped at US/Republic of Korea (ROK) military training sites near the demilitarized zone (DMZ) of Paju, Pocheon, and Yeoncheon, Gyeonggi Province from December 2004 to December 2009. Small mammals were examined for their intestinal nematodes by necropsy. A total of 1,617 rodents (100%) and 91 (100%) soricomorphs were infected with at least 1 nematode species, including Nippostrongylus brasiliensis, Heligmosomoides polygyrus, Syphacia obvelata, Heterakis spumosa, Protospirura muris, Capillaria spp., Trichuris muris, Rictularia affinis, and an unidentified species. N. brasiliensis was the most common species infecting small mammals (1,060; 62.1%) followed by H. polygyrus (617; 36.1%), S. obvelata (370; 21.7%), H. spumosa (314; 18.4%), P. muris (123; 7.2%), and Capillaria spp. (59; 3.5%). Low infection rates (0.1-0.8%) were observed for T. muris, R. affinis, and an unidentified species. The number of recovered worms was highest for N. brasiliensis (21,623 worms; mean 20.4 worms/infected specimen) followed by S. obvelata (9,235; 25.0 worms), H. polygyrus (4,122; 6.7 worms), and H. spumosa (1,160; 3.7 worms). A. agrarius demonstrated the highest prevalence for N. brasiliensis (70.9%), followed by M. minutus (50.0%), T. triton (33.3%), M. fortis (28.1%), M. musculus (15.6%), C. lasiura (13.2%), and M. regulus (0%). This is the first report of nematode infections in small mammals captured near the DMZ in ROK.     

Detection of Foodborne Bacterial Pathogens from Individual Filth Flies

There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a particular foodborne illness outbreak has been challenging. This is because insects are not being aseptically collected as part of an environmental sampling program during foodborne outbreak investigations and because there is not a standardized method to detect foodborne bacteria from individual insects. To take a step towards solving this problem, we adapted a protocol from a commercially available PCR-based system that detects foodborne pathogens from food and environmental samples, to detect foodborne pathogens from individual flies.Using this standardized protocol, we surveyed 100 wild-caught flies for the presence of Cronobacter spp., Salmonella enterica, and Listeria monocytogenes and demonstrated that it was possible to detect and further isolate these pathogens from the body surface and the alimentary canal of a single fly. Twenty-two percent of the alimentary canals and 8% of the body surfaces from collected wild flies were positive for at least one of the three foodborne pathogens. The prevalence of Cronobacter spp. on either body part of the flies was statistically higher (19%) than the prevalence of S. enterica (7%) and L.monocytogenes (4%). No false positives were observed when detecting S. enterica and L. monocytogenes using this PCR-based system because pure bacterial cultures were obtained from all PCR-positive results. However, pure Cronobacter colonies were not obtained from about 50% of PCR-positive samples, suggesting that the PCR-based detection system for this pathogen cross-reacts with other Enterobacteriaceae present among the highly complex microbiota carried by wild flies. The standardized protocol presented here will allow laboratories to detect bacterial foodborne pathogens from aseptically collected insects, thereby giving public health officials another line of evidence to find out how the food was contaminated when performing foodborne outbreak investigations.     

Endosymbionts of Acanthamoeba Isolated from Domestic Tap Water in Korea

In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the α-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.     

Molecular Detection of Giardia intestinalis from Stray Dogs in Animal Shelters of Gyeongsangbuk-do (Province) and Daejeon,Korea

Giardia is a major public health concern and considered as reemerging in industrialized countries. The present study investigated the prevalence of giardiosis in 202 sheltered dogs using PCR. The infection rate was 33.2% (67/202); Gyeongsangbuk-do and Daejeon showed 25.7% (39/152, P<0.0001) and 56% (28/50), respectively. The prevalence of infected female dogs (46.7%, P<0.001) was higher than in male dogs (21.8%). A higher prevalence (43.5%, P<0.0001) was observed in mixed breed dogs than purebred (14.1%). Although most of the fecal samples collected were from dogs of ≥1 year of age which showed only 27.4% positive rate, 61.8% (P<0.001) of the total samples collected from young animals (<1 year of age) were positive for G. intestinalis. A significantly higher prevalence in symptomatic dogs (60.8%, P<0.0001) was observed than in asymptomatic dogs (23.8%). Furthermore, the analysis of nucleotide sequences of the samples revealed that G. intestinalis Assemblages A and C were found in the feces of dogs from Gyeongsangbuk-do and Daejeon. Since G. intestinalis Assemblage A has been known to infect humans, our results suggest that dogs can act as an important reservoir of giardiosis in Korea. Hence, hygienic management should be given to prevent possible transmission to humans.     

舟山群岛春秋季獐栖息地的生态特征

2008 年3 月和2008 年11 月,在舟山群岛以獐的足迹、粪便和卧迹等新鲜活动痕迹为依据,对獐春、秋季栖息地利用特征进行研究。共设置样方420 个,对样方内生境类型、乔木盖度、灌木盖度、草本盖度、坡位、坡度、坡向、海拔、人为干扰距离和水源距离等10 个生态因子进行测量评估。结果发现,獐春、秋两季的栖息地利用特征是(1)隐蔽和食物因子:春、秋季利用阔叶林、农田和山坡地,秋季对灌木林也有较高的利用率,而对农田的利用率下降;春、秋季乔木盖度、灌木盖度和草本盖度较低处(≤50%) 利用率较高,但春季对乔木盖度较高处(> 50% )也有较高的利用率; (2) 地形因子:春季对坡度较缓、中下坡位、海拔较低处(< 100 m)利用率较高;秋季对坡度较缓、中下坡位、海拔较低(<100 m)的南坡利用率较高; (3) 水源因子:春季利用水源距离较近(< 200 m) 的区域;秋季对水源距离较近(< 200 m)和较远(>600 m) 区域的利用率均较高; (4) 干扰因子:春季主要利用距离人为干扰近处(< 100 m),秋季主要利用距离人为干扰远(> 200 m)处。逐步判别分析显示,春、秋季獐栖息地利用特征存在显著差异,乔木盖度、坡度、坡向、海拔和人为干扰5 个因子是主要的区分因子。舟山群岛人为干扰剧烈、次生乔木和灌木较为发达以及草本植物不发达等一系列特点,造成了舟山群岛獐特殊的栖息地现状。本研究将对制订适合于海岛的动物栖息地保护对策提供理论依据,也为了解海岛生境下獐不同季节的生存状况提供科学依据。     

Anisakis pegreffii Larvae in Sea Eels (Astroconger myriaster) from the South Sea,Republic of Korea

Anisakis simplex sensu stricto (s.s.), Anisakis pegreffii, Anisakis berlandi (=A. simplex sp. C), and Anisakis typica are the 4 major species of Anisakis type I larvae. In the Republic of Korea (Korea), A. pegreffii, A. berlandi, and A. typica larvae in fish hosts has seldom been documented. In this study, molecular analysis was performed on Anisakis larvae from the sea eels (Astroconger myriaster), the major source of human anisakiasis in Korea, collected from Tongyeong City, a southern coastal area of Korea. All 20 sea eels examined were infected with Anisakis type I larvae (160 larvae; 8 per fish). Their species were analyzed using PCR-RFLP patterns and nucleotide sequences of internal transcribed spacers (ITS1, 5.8 subunit gene, and ITS2) and mitochondrial cytochrome c oxidase 2 (cox2). Most (86.8%; 112/129) of the Anisakis type I larvae were A. pegreffii, and 7.8% (10/129) were A. typica. The remaining 5.4% (7/129) was not identified. Thus, A. pegreffii is the major species of anisakid larvae in sea eels of the southern coast of Korea.     

Echinostome Infections in the Striped-Field Mouse, Apodemus agrarius, and the Ussuri White-Toothed Shrew, Crocidura lasiura, Caught Near the Demilitarized Zone, Gyeonggi-do (Province), Republic of Korea

A total of 1,498 small mammals (rodents and insectivores), including Apodemus agrarius (n = 1,366), Crocidura lasiura (54), Mus musculus (32), Micronytus fortis (28), Eothenomys regulus (9), Micronys minutes (6), and Cricetulus triton (3), were live-trapped in Gyeonggi-do (Province) (Paju-si, Pocheon-gun, and Yeoncheon-gun) near the demilitarized zone (DMZ) from December 2004 to September 2005. A. agrarius was found to be infected with 3 species of echinostomes (Echinostoma hortense, Echinostoma cinetorchis, and Euparyphium murinum), while C. lasiura was infected with 1 species (Echinochasmus japonicas) of echinostome. Other mammals were free from echinostome infections. Total 16 E. hortense were detected in 7 (0.5%) mice, 9 E. cinetorchis from 5 (0.4%), and 3 E. murinum from 2 (0.1%) out of 1.366 A. agrarius examined. E. japonicus was found only in 1 (1.9%; total 3 specimens) C. lasiura. These results demonstrate that A. agrarius and C. lasiura, inhabiting near the DMZ of Gyeonggi-do serve as the natural definitive hosts for several species of echinostomes, although their infection rates are low. This is the first record of natural infections of A. agrarius with E. cinetorchis and C. lasiura with E. japonicus in the Republic of Korea.     

Ornithodoros sawaii (Ixodida: Argasidae) Larvae Collected from Hydrobates monorhis on Sogugul and Gaerin Islands,Jeollanam-do (Province), Republic of Korea

The 65th Medical Brigade and Public Health Command District-Korea, in collaboration with the Migratory Bird Research Center, National Park Research Institute, conducted migratory bird tick surveillance at Sogugul and Gaerin Islands (small rocky bird nesting sites), Jeollanam-do (Province), Republic of Korea (ROK), on 30 July and 1 August 2009. Breeding seabirds captured by hands in their nesting burrows were banded, identified to species, and carefully examined for ticks during the nesting season. A total of 9 Ornithodoros sawaii larvae were removed from 4 adult Hydrobates monorhis (Swinhoe’s storm petrel). The identification of the larvae of O. sawaii collected from migratory seabirds were molecularly confirmed using mitochondrial 16S rDNA primer sets.