Epidemiological survey and genetic diversity of Bartonella in fleas collected from rodents in Fujian Province, Southeast China.

BACKGROUND: Fleas, considered to be the main transmission vectors of Bartonella, are highly prevalent and show great diversity. To date, no investigations have focused on Bartonella vectors in Southeast China. The aim of this study was to investigate the epidemiological and molecular characteristics of Bartonella in fleas in Southeast China. METHODS: From 2016 to 2022, flea samples (n = 1119) were collected from 863 rodent individuals in seven inland and coastal cities in Southeast China. Flea species, region, gender, host species and habitat were recorded. The DNA samples from each individual flea were screened by real-time PCR for the Bartonella ssrA gene. All positive samples were confirmed by PCR based on the presence of the gltA gene and sequenced. The factors associated with Bartonella infection were analyzed by the Chi-square test and Fisher's exact test. ANOVA and the t-test were used to compare Bartonella DNA load. RESULTS: Bartonella DNA was detected in 26.2% (293/1119) of the flea samples, including in 27.1% (284/1047) of Xenopsylla cheopis samples, 13.2% (5/38) of Monopsyllus anisus samples, 8.3% (2/24) of Leptopsylla segnis samples and 20.0% (2/10) of other fleas (Nosopsyllus nicanus, Ctenocephalides felis, Stivalius klossi bispiniformis and Neopsylla dispar fukienensis). There was a significant difference in the prevalence of Bartonella among flea species, sex, hosts, regions and habitats. Five species of Bartonella fleas were identified based on sequencing and phylogenetic analyses targeting the gltA gene: B. tribocorum, B. queenslandensis, B. elizabethae, B. rochalimae and B. coopersplainsensis. CONCLUSIONS: There is a high prevalence and diversity of Bartonella infection in the seven species of fleas collected in Southeast China. The detection of zoonotic Bartonella species in this study, including B. tribocorum, B. elizabethae and B. rochalimae, raises public health concerns.

Prevalence and phylogenetic analysis of Babesia parasites in reservoir host species in Fujian province, Southeast China.

Babesiosis is a tick-borne disease that mainly affects small mammals and has been reported in at least five provinces in China. However, the host range and geographical distribution of the parasite in Fujian province are unclear. Therefore, we investigated the prevalence and genetic characteristics of Babesia in Fujian province, Southeast China, between 2015 and 2020. Rodent blood samples were collected from 26 different surveillance sites across Fujian province. Genomic DNA was extracted to screen for Babesia infection using polymerase chain reaction based on 18S rRNA. DNA samples from 316 domestic goats, 85 water buffalo, 56 domestic dogs and 18 domestic pigs were examined. The prevalence of Babesia was statistically analysed using the Chi-square test or Fisher's exact test. Babesia infections were detected in 3.96% (43/1,087; 95%CI: 2.80%, 5.12%) of rodents and 1.26% (6/475; 95%CI: 0.26%, 2.26%) of other mammals. Multivariate logistic regression analysis revealed that irrigated cropland, shrubs and forests were risk factors for Babesia microti infections. The infection rates among domestic pigs, dogs and goats were 5.56%, 1.79% and 1.27%, respectively, with no infection found in water buffalo. The 18S rRNA gene sequencing revealed that rodents were infected with Babesia (sensu lato), whereas other mammals were infected with Babesia (sensu stricto). The geographical distribution and phylogenetic relationship of Babesia was determined in Southeast China. Mammals, particularly wild rodents, maybe the main natural hosts of Babesia in Fujian. Our findings provide a foundation for public health officials to develop prevention and control measures for Babesia.

Serological and molecular characteristics of pathogenic Leptospira in rodent populations in Fujian Province, China, 2018-2020.

BACKGROUND: Leptospirosis is a significant emerging infectious disease worldwide. Rodents are considered to be the most critical hosts of Leptospira spp. Fujian Province is a region highly endemic for leptospirosis in China. However, the genetic diversity of leptospires circulating among rodents in Fujian is limited. RESULTS: The carrier status of rodents for Leptospira spp. was investigated by culture and serological detection in Fujian during 2018-2020. A total of 710 rodents, including 11 species, were trapped, with Rattus losea being the dominant trapped species (50.56%). Fourteen pathogenic Leptospira strains were obtained. Seven L. borgpetersenii serogroup Javanica strains belonging to ST143, 4 L. interrogans serogroup Icterohaemorrhagiae strains belonging to ST1 and ST17, 2 L. interrogans serogroup Bataviae strains belonging to ST96 and ST333, and 1 L. interrogans serogroup Pyrogenes strains belonging to ST332 were identified using 16S rDNA gene sequencing, microscopic agglutination test (MAT) and Multilocus sequence typing (MLST). L. borgpetersenii serogroup Javanica belonging to ST143 was the dominant type (50.00%). A total of 387 rodent serum samples were tested by MAT. Serum were considered positive for seroreactivity at a titer ≥ 1:160 against at least one serovar. A total of 90 (23.26%) serum samples tested positive, and four serogroups were identified, with Javanica being the dominant serogroup (87.78%), which was similar to the dominant serogroup isolated from rodents. This study demonstrates a high prevalence of leptospirosis in rodents and public health education among high-risk workers is highly recommended. CONCLUSIONS: R. losea was the dominant trapped rodent, and L. borgpetersenii serogroup Javanica ST143 was widely distributed among rodents in Fujian from 2018 to 2020. Despite the low number of isolates obtained from rodents, this study suggests that continuous epidemiological surveillance of the aetiological characteristics of pathogenic Leptospira in wild animal reservoirs may help reduce the possible risk of disease transmission.

Epidemiological characteristics and genetic diversity of Bartonella species in rodents from southeastern China.

Rodents are the primary hosts of Bartonella species and carry more than 22 Bartonella species. However, the information on epidemiological characteristics and genetic diversity of Bartonella species in rodents in southeastern China is limited. From 2015 to 2020, 1,137 rodents were captured. Bartonella-positive DNA was detected in 14.9% (169/1,137) of rodents by PCR for both the ssrA and gltA genes. A highest Bartonella prevalence was detected in Apodemus agrarius (33.5%) and lowest in B. indica (1.8%). The probability of Bartonella infection in summer (20.1%) was higher than in spring (14.6%; p = .011, OR = 1.756). Sequencing and phylogenetic analysis revealed that nine known Bartonella species were identified in rodents, including B. tribocorum, B. grahamii, B. rattimassiliensis, B. queenslandensis, B. elizabethae, B. phoceensis, B. coopersplainsensis, B. japonica and B. rochalimae. In our study, Bartonella species exhibited a strong association with their hosts. Zoonotic B. tribocorum, B. grahamii, B. elizabethae and B. rochalimae were found in synanthropic rodent species in southeastern China, which pose a potential threat to the public health. To prevent the spread of zoonotic Bartonella species to humans, preventive and control measures should be implemented, and more research is needed to confirm the pathogen's association with human disease.

[Investigation and genetic identification on Babesia infection in rodents in some areas of Fujian Province].

Objective: To explore the status of Babesia infection in rodents and the genetic characteristics of Babesia spp. in Fujian Province. Methods: Rodents were captured by the night trapping method in Shaowu, Qingliu, Shunchang, Yong'an, Changle and Youxi during 2014-2015. The rodent species was identified, and information on the time and place of capture, species and sex of rodents was recorded. Blood samples was collected, in which the fragment of 18S rRNA gene of Babesia spp. was amplified by PCR. The PCR products were sequenced and the phylogenetic tree was constructed for homology analysis. Data on positive rate were analyzed with Chi-square or Fisher exact test. Results: Two hundred and nine rats were captured, comprising of 71 domestic and 138 wild rats. The overall positive rate was 9.6%(20/209). The positive rate in domestic rats was 2.8%(2/71), including one Rattus norvebicus and one Rattus flavipectus. The positive rate in wild rats was 13.0%(18/138), including 13 Bandicota indica, one Rattus losea, 2 Rattus confucianus and 2 Rattus fulvescens. The positive rate was significantly higher in wild rats than in domestic rats (P < 0.05). The Youxi region had the highest positive rate(14.9%,13/87), followed by Yong'an(13.6%, 3/22), and no positive rat was found in Qingliu. The positive rate in the male rats was 7.9%(9/114), and that in the females was 11.6%(11/95). The positive rate was highest in adult rats (10.4%,18/173), followed by young ones (6.3%,2/32). No positive rat was found in old rats. There was no significant difference in positive rate among different regions, between male and female rats, or among different ages (P > 0.05). The sequences of PCR products had a 100% homology. The BLAST results revealed the species to be Babesia microti. The phylogenetic tree showed that the sample sequence was the most homologous with Babesia microti from Zhejiang Province(GenBank Accession No: JQ609305). Conclusion: There occurs Babesia microti infection in rats in part areas of Fujian Province. The positive rate was higher in wild rats than in domestic rats.

The effect of temperature on the extrinsic incubation period and infection rate of dengue virus serotype 2 infection in Aedes albopictus.

Dengue fever is an acute mosquito-borne viral disease caused by dengue virus (DENV). Temperature may affect the efficiency of the mosquito vectors in spreading DENV. Aedes albopictus mosquitoes were infected orally with a DENV2 suspension and incubated at different temperatures. Subsequently, DENV2 antigen was collected from salivary gland and thorax-abdomen samples on different days postinfection and tested using an immunofluorescence assay to determine the extrinsic incubation period and infection rate. As the temperature increased, the extrinsic DENV2 incubation period in Ae. albopictus gradually shortened, and infection rates showed a tendency to initially increase, followed by a subsequent decrease.

[DNA amplification of Plasmodium vivax parasites from Giemsa-stained blood smears].

OBJECTIVE: To develop methods of extracting DNA from malaria parasites on Giemsa-stained blood smears. METHODS: Improved Na2HPO4 method and Chelex-100 ion-exchange technique were used to extract DNA from Giemsa-stained or unstained blood smears. Nested PCR was employed for amplification and identification of allelotype in the Plasmodium vivax merozoite surface protein-1 (PvMSP-1). RESULTS: Target DNA bands appeared in all samples of unstained thick blood smears, while no DNA bands were visible in the fixed and stained thin smears. Both methods identified PvMSP-1 alleles from smears with parasitemia of > or = 0.01%. CONCLUSION: It is feasible to identify PvMSP-1 alleles from Giemsa-stained blood smear.