Animals ; Disease Reservoirs ; microbiology ; Genome, Bacterial ; Geography ; Marmota ; microbiology ; Plague ; epidemiology ; microbiology ; Polymorphism, Genetic ; Yersinia pestis ; genetics

The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.
Acanthamoeba castellanii/*microbiology ; Animals ; Disease Reservoirs/*microbiology ; Disease Vectors ; Escherichia coli/growth & development/pathogenicity/*physiology ; Oocysts/microbiology ; Symbiosis/*physiology ; Trophozoites/microbiology

OBJECTIVETo study the pathogenic ecology characteristics of plague in Qinghai plateau.

METHODSApplied molecular biology techniques, conventional technologies and geographic information system (GIS) to study phenotypic traits, plasmid spectrum, genotype, infected host and media spectrum etc.of 952 Yersinia pestis strains in Qinghai plateau plague foci, which were separated from different host and media in different regions during 1954 to 2012.

RESULTSThe ecotypes of these strains were Qingzang plateau (91.49%, 871/952),Qilian mountain (6.41%, 61/952) and Microtus fuscus (1.26%, 12/952).83.6% (796/952) of these strains contained all the 4 virulence factors (Fr1, Pesticin1,Virulence antigen, and Pigmentation), 93.26% (367/392) were velogenic strains confirmed by virulence test.725 Yersinia pestis strains were separated from Qinghai plateau plague foci carried 9 kinds of plasmid, among which 713 strains from Marmot himalayan plague foci carried 9 kinds of plasmid, the Mr were 6×10(6), 7×10(6), 23×10(6), 27×10(6), 30×10(6), 45×10(6), 52×10(6), 65×10(6) and 92×10(6) respectively. 12 Yersinia pestis strains were separated from Microtus fuscus plague foci carried only 3 kinds of plasmid, the Mr were 6×10(6), 45×10(6), 65×10(6). Meanwhile, the strains carrying large plasmid (52×10(6), 65×10(6) and 92×10(6)) were only distributed in particular geographical location, which had the category property. The research also confirmed that 841 Yersinia pestis strains from two kinds of plague foci in Qinghai plateau had 11 genomovars. The strains of Marmot himalayan plague foci were given priority to genomovar 5 and 8, amounted to 611 strains, genomovar 8 accounted for 56.00% (471/841), genomovar 5 accounted for 23.07% (194/841). Besides, 3 new genomovars, including new 1(62 strains), new 2(52 strains), new 3(48 strains) were newly founded, and 12 strains of Microtus fuscus plague foci were genomovar 14.

CONCLUSIONThe main host and media of Qinghai plateau plague foci directly affected the spatial distribution regularities of plague epidemic and the pathogens characteristics, meanwhile the polymorphism of plague ecological geographic landscape leds to the complexity of Yersinia pestis' genotype.

Animals ; Arvicolinae ; microbiology ; China ; epidemiology ; Disease Reservoirs ; microbiology ; Ecology ; Genotype ; Marmota ; microbiology ; Plague ; epidemiology ; microbiology ; Virulence ; genetics ; Yersinia pestis ; genetics ; pathogenicity

OBJECTIVETo investigate Bartonella infections in small mammalian reservoir hosts from different environments and types of climate in Yunnan.

METHODSFemoral blood samples were collected from the anesthetic captured animals from five counties including three types of climate. All isolates were grown on brain and heart infusion agar plates containing 5% defibrinated rabbit blood. The agar plates were incubated at 35 degrees C in a humidified with 5% CO2 environment for at least 4 weeks. Bartonella-like isolates were confirmed by the polymerase chain reaction and visualizing the target gene fragment by gel electrophoresis.

RESULTSBartonella species were isolated from 69 of 176 small animals including 4 species of 3 genera from 4 counties and the total prevalence in rodents was 39.2%. The maximal prevalence was 42.0% of Rattus tanezumi flavipectus usually inhabiting indoors and courtyard and contacting closely to human. Moreover, Bartonella isolates were obtained from Rattus noruegicus, Eothenomys miletus and Mus pahari. Life environments of captured animals involved indoors, courtyard, brush and forest in mountain.

CONCLUSIONThe finding in this study suggested the characteristic of diversity of Bartonella infections in rodent hosts in southern China included Bartonella species parasiting in a wide range of animal hosts in different environments as well as climate types. Further investigations were needed in different areas in China to confirm more mammalian reservoir hosts with Bartonella infections.

Animals ; Bartonella ; classification ; genetics ; isolation & purification ; Bartonella Infections ; epidemiology ; veterinary ; China ; epidemiology ; Disease Reservoirs ; Mice ; Rats ; microbiology ; Rodent Diseases ; microbiology ; Rodentia ; microbiology ; Species Specificity

OBJECTIVETo study the increasing incidence and the characteristics of Tsutsugamushi disease in the areas of Nan Peng Lie islands, Nan Ao island, Wan Shan archipelago, Nao Zhou island and Lei Zhou peninsula, located in the southern part of China and to develop strategies for preventive measures.

METHODSBoth epidemiological investigation, isolation and gene identification of Orientia tsutsugamushi, as well as pilot preventive measures were carried out.

RESULTSThese islands belonged to the epidemic area of south subtropical zone of Tsutsugamushi disease. The main host was Rattus norvegicu and the overall rates of infection on Orientia tsutsugamushi were 22.78%-33.75%. The main biological vector was Leptotrombidium (Leptotrombidium) deliens and the rates of infection on Orientia tsutsugamushi were 40.00%-75.00%. 25 strains of Orientia tsutsugamushi had been isolated from Rattus norvegicu and Leptotrombidium (Leptotrombidium) deliens. Results showed that the isolated strains of Orientia tsutsugamushi were 15 Karp, 8 Kato, 2 Yonchon. Results from serological studies showed that the positive rate of anti-Orientia tsutsugamushi antibodies was high, in both residents and soldiers stationed in these islands. On these islands, rats and biological vectors were killed. Results showed that these measures had positive impact in reducing the incidence.

CONCLUSIONIslands from the southern part of the country belonged to the epidemic area of Tsutsugamushi disease. People visiting this areas should be under protection.

Animals ; Antibodies, Bacterial ; blood ; Bacterial Typing Techniques ; China ; epidemiology ; Disease Outbreaks ; Disease Reservoirs ; microbiology ; Geography ; Humans ; Incidence ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Rats ; Scrub Typhus ; epidemiology ; Trombiculidae ; microbiology

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.
Anaplasma/genetics/*physiology ; Animals ; Arachnid Vectors/microbiology ; Bartonella/genetics/*physiology ; Chaperonin 60/genetics ; Deer/parasitology ; Disease Reservoirs/veterinary ; RNA, Ribosomal, 16S/genetics ; Republic of Korea/epidemiology ; Ticks/*microbiology

Comprehensive quarterly serosurveillance on scrub typhus in small mammals collected from military training sites located near the Demilitarized Zone (DMZ), northern Gyeonggi-do (Province), ROK was conducted to determine the potential rodent-borne and associated ectoparasite disease risks to military personnel. A total of 1,196 rodents and insectivores representing 8 species, Apodemus agrarius (87.3%, n = 1,044), Mus musculus (5.4%, n = 65), Crocidura lasiura (3.3%, n = 40), Microtus fortis (2.6%, n = 31), Micromys minutus (0.3%, n = 4), Tscherskia triton (0.3%, n = 4), Rattus norvegicus (0.3%, n = 4), and Myodes regulus (0.3%, n = 4) were assayed for the presence of antibodies to Orientia tsutsugamushi. O. tsutsugamushi antibodies were detected in 6 of 8 species and seroprevalence determined; A. agrarius (45.6%), M. musculus (23.1%), M. fortis (48.4%), M. minutus (50.0%), T. triton (50.0%), and R. norvegicus (25.0%). A total of 31,184 chigger mites collected from 508 rodents and insectivores were slide-mounted and 10 species belonging to 4 genera were identified. Leptotrombidium pallidum (53.4%) was the most frequently collected, followed by L. palpale (15.7%), Neotrombicula tamiyai (14.3%), L. orientale (10.7%), L. zetum (3.1%), Walchia fragilis (2.1%), and L. gemiticulum (0.8%), while the remaining 3 species, L. subintermedium, N. gardellai, and Euschoengastia koreaensis were rarely observed (prevalence < 10%). In contrast to previous surveys, higher chigger indices of the primary scrub typhus vectors, L. pallidum (165.4), L. orientale (45.0), and L. palpale (21.4), were observed during the spring season.
Animals ; Antibodies, Bacterial/immunology ; Arachnid Vectors/classification/*microbiology ; Disease Reservoirs/classification/microbiology/*parasitology ; Humans ; Military Facilities ; Orientia tsutsugamushi/*immunology/isolation & purification ; Republic of Korea ; Rodentia/classification/*immunology/microbiology/*parasitology ; Scrub Typhus/microbiology/*transmission/*veterinary ; Seroepidemiologic Studies ; Trombiculidae/classification/*microbiology

Blood, saliva, and nail samples were collected from 54 dogs and 151 cats and analyzed for the presence of Bartonella henselae with a novel nested polymerase chain reaction (PCR) method. Bartonella (B.) henselae was detected in feral cat blood (41.8%), saliva (44.1%), and nail (42.7%) samples. B. henselae was also detected in pet cat blood (33.3%), saliva (43.5%), and nail (29.5%) samples and in pet dog blood (16.6%), saliva (18.5%), and nail (29.6%) samples. Nine samples were infected with B. clarridgeiae and 2 were co-infected with B. henselae and B. clarridgeiae of blood samples of dogs. This report is the first to investigate the prevalence of B. henselae and B. clarridgeiae in dogs and cats in Korea, and suggests that dogs and cats may serve as potential Bartonella reservoirs.
Animals ; Bartonella/*classification ; Bartonella Infections/blood/epidemiology/microbiology/*veterinary ; Cat Diseases/blood/epidemiology/*microbiology ; Cats ; Disease Reservoirs/veterinary ; Dog Diseases/epidemiology/*microbiology ; Dogs ; Hoof and Claw/microbiology ; Korea/epidemiology ; Prevalence ; Saliva/microbiology

OBJECTIVETo study the correlation between Vibrio cholerae strains isolated from natural enviroment and fishery products and the source of infection during V. cholerae outbreaks.

METHODSCholera toxin gene was detected by polymerase chain reaction (PCR) amplification. Pulsed-field gel electrophoresis (PFGE) was used to subtype the isolates. Results of PFGE were analyzed and clustered by BioNumerics software (Version 4.0).

RESULTSDuring the outbreaks, a total number of thirty O139 V. cholerae related serogroups were collected from patients, carriers, sewage and fishery products were identified and proved to be toxigenic. They could be clustered into four PFGE patterns when digested by Not I. These two V. cholerae outbreaks were caused by the same source of infection because of the following reasons: (1) PFGE patterns of the predominant strains isolated from two outbreaks were identical; (2) they were identical to the PFGE patterns of the strains isolated from the green turtle and rana catesbiana which were bought from the same wholesale store.

CONCLUSIONGreen turtle and rana catesbiana that were contaminated by toxigenic O139 V. cholerae strains seemed to be the source of infection causing the O139 V. cholerae outbreaks in Jiangxi province. Rapid laboratory surveillance and epidemiologic investigation were important in identifying the source of infection during the outbreaks of V. cholera.

Animals ; China ; epidemiology ; Cholera ; epidemiology ; Cluster Analysis ; Disease Outbreaks ; Disease Reservoirs ; microbiology ; Electrophoresis, Gel, Pulsed-Field ; methods ; Fisheries ; Humans ; Ranidae ; microbiology ; Sewage ; microbiology ; Turtles ; microbiology ; Vibrio cholerae O139 ; isolation & purification

OBJECTIVETo isolate and identify Bartonella strains from native dogs in Shandong province in China.

METHODSEDTA-anticoagulated blood samples were collected from 71 native dogs in Yanggu county of Shandong province in March 2005. All isolates were grown on brain heart infusion agar plates containing 5% defibrinated rabbit blood. The agar plates were incubated at 37 degrees C in a humidified with 5% CO2 environment for 4 weeks or longer. All Bartonella-like isolates were examined by routine Gram and Giménez staining and then followed by polymerase chain reaction (PCR) and PCR-RFLP analysis for identification and differentiation of the isolates. Sequencing 16S rRNA, citrate synthase (gltA) gene and 16S-23S rRNA ITS were carried out and sequential similarities were calculated using the DNASTAR5 software package. The phylogenetic tree was inferred from each bootstrap sample, using the neighbor-joining methods as executed in the MEGA 3.1 software. The translation from DNA to protein were determined by DNASIS 2.5.

RESULTSThe two Bartonella-like organisms (strains Q52SHD and Q64SHD) were isolated from the blood of 71 dogs. Light microscopic examination of the Gram and Giménez-stained micro-organisms showed small, short and slightly curved pleomorphic gram-negative bacilli. Amplified products of the three pairs of Bartonella genus-specific primers carried the same size as the predicted of those Bartonella species. Data from PCR-RFLP analysis showed that the two strains that having the same profiles were all different from the B. henselae type strain-16S rRNA, gltA and 16S-23S rRNA ITS sequences from the two isolates were 100.0%, 99.7% and 97.2% homologous to B. vinsonii berkhoffii.

CONCLUSIONSBased on these findings, the two isolates Q52SHD and Q64SHD were demonstrated as B. vinsonii berkhoffii. To our knowledge, this was the first report on the presence of Bartonella infection in native dogs from China, which constituted a large reservoir of Bartonella species in this country.

Animals ; Bartonella ; classification ; genetics ; isolation & purification ; Bartonella Infections ; veterinary ; Disease Reservoirs ; Dogs ; microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; Rabbits