Association of Bartonella with the fleas (Siphonaptera) of rodents and bats using molecular techniques.

Bartonella spp. are putatively vector-borne bacterial agents of humans and animals. Fleas have been incriminated as vectors of Bartonella spp. and are suspected of transmitting Bartonella of rodents and bats, but some of these Bartonella spp. have not yet been directly detected in wild caught fleas. We report the molecular detection of Bartonella tribocorum, Bartonella vinsonii subsp. vinsonii, and two novel genotypes of Bartonella from the fleas Xenopsylla cheopis, Ctenophthalmus pseudagyrtes, Sternopsylla texanus, or Orchopeas howardi.     

中国云南人间鼠疫流行区蚤类宿主特异性及宿主选择(英文)

应用生态学中的生态位宽度及生态位重叠研究方法,对中国云南人间鼠疫流行区１１种蚤的宿主特异性及宿主选择进行了研究。宿主特异性用Ｌｅｖｉｎｓ生态位宽度进行测定。宿主选择用夹角余弦生态位重叠进行测定。结果表明：长形病蚤普洱亚种及印鼠客蚤的生态位宽度最窄、宿主特异性最高,近端远棒蚤二刺亚种及斯氏新蚤滇川亚种的生态位最宽、宿主特异性最低。印鼠客蚤（云南人间鼠疫流行区的主要鼠疫媒介）的优势宿主是黄胸鼠。印鼠客蚤较高的宿主特异性意味着该蚤可能主要在其优势宿主的不同个体间保存或传播鼠疫病原体。短突栉眼蚤及端凹栉眼蚤在宿主选择上较接近,而其它蚤种在宿主选择上差异较大。     

中国云南部分人间鼠疫流行区蚤类区系调查(英文)

归纳了中国云南 13个人间鼠疫流行区的调查资料 ,对调查疫区的蚤类区系进行了研究。总计捕获12 0 77只小兽 ,隶属啮齿目、食虫目及攀目 3个目中的 9科、2 9属、4 7种。从小兽体表共采获 9369只蚤 ,经分类鉴定 ,隶属 5科、18属、33种。 33种蚤及 4 7种小兽宿主均按其分类阶元详细列于文末。结果表明 ,山区蚤及小兽宿主的种数明显多于坝区。坝区农耕地的优势种相对简单 ,优势种地位突出 ,黄胸鼠及印鼠客蚤分别是最重要的宿主及蚤种 (构成比分别为 83 2 7%和 75 32 % )。山区的优势种相对较复杂 ,优势种的种类较多 ,但其构成比较低 ( 10 96%～ 4 7 95% )。黄胸鼠及绒鼠为山区地带的两种优势宿主 ,缓慢细蚤、端凹栉眼蚤、印鼠客蚤、偏远古蚤及短突栉眼蚤为山区地带的 5种优势蚤种。多数蚤种可寄生两种以上的小兽宿主 ,但其所寄生的主要宿主并不多。结果提示 ,作为疫区主要媒介的印鼠客蚤及其所对应的主要寄生宿主 (黄胸鼠 )在坝区突出的优势种地位 ,似可解释近年疫区的鼠疫病人主要出现在坝区的原因     

Molecular detection and identification of Bartonella species in Xenopsylla cheopis fleas (Siphonaptera: Pulicidae) collected from Rattus norvegicus rats in Los Angeles, California

Of 200 individual Xenopsylla cheopis fleas removed from Rattus norvegicus rats trapped in downtown Los Angeles, CA, 190 (95%) were positive for the presence of Bartonella DNA. Ninety-one amplicons were sequenced: Bartonella rochalimae-like DNA was detected in 66 examined fleas, and Bartonella tribocorum-like DNA was identified in 25 fleas. The data obtained from this study demonstrate an extremely high prevalence of Bartonella DNA in rat-associated fleas.     

Investigation of Bartonella acquisition and transmission in Xenopsylla ramesis fleas (Siphonaptera: Pulicidae)

Bartonella are emerging and re-emerging pathogens affecting humans and a wide variety of animals including rodents. Horizontal transmission of Bartonella species by different hematophagous vectors is well acknowledged but vertical transmission (from mother to offspring) is questionable and was never explored in fleas. The aim of this study was to investigate whether the rodent flea, Xenopsylla ramesis, can acquire native Bartonella from wild rodents and transmit it transovarially. For this aim, Bartonella-free laboratory-reared X. ramesis fleas were placed on six naturally Bartonella-infected rodents and six species-matched Bartonella-negative rodents (three Meriones crassus jirds, two Gerbillus nanus gerbils and one Gerbillus dasyurus gerbil) for 7 days, 12-14h per day. The fleas that were placed on the Bartonella-positive rodents acquired four different Bartonella genotypes. Eggs and larvae laid and developed, respectively, by fleas from both rodent groups were collected daily for 7 days and molecularly screened for Bartonella. All eggs and larvae from both groups were found to be negative for Bartonella DNA. Interestingly, two of five gut voids regurgitated by Bartonella-positive fleas contained Bartonella DNA. The naturally infected rodents remained persistently infected with Bartonella for at least 89 days suggesting their capability to serve as competent reservoirs for Bartonella species. The findings in this study indicate that X. ramesis fleas can acquire several Bartonella strains from wild rodents but cannot transmit Bartonella transovarially.     

云南省玉龙鼠疫疫源地野外鼠形动物寄生蚤丰盛度影响因素分析

【目的】分析云南省玉龙鼠疫疫源地野外鼠形动物寄生蚤丰盛度的影响因素。【方法】选取云南省玉龙鼠疫疫源地3个海拔区域,按4个季节进行野外捕鼠,捕获的鼠形动物用梳检法收集体表寄生蚤并在显微镜下分类鉴定。通过实际测量和实地观察相结合的方式收集潜在影响鼠形动物寄生蚤丰盛度的因素包括鼠形动物特征变量指标(如种类、年龄、性别、体长、体重)、环境和气象因子(如海拔、季节)等数据。采用EpiData 3.02软件建立数据库,在R软件下使用跨栏负二项分布回归分析鼠形动物寄生蚤丰盛度的影响因素。【结果】从捕获的884只鼠形动物中检获寄生蚤9种484头,以特新蚤指名亚种、方叶栉眼蚤、无值大锥蚤、云南栉眼蚤为主(86.16%)。回归分析显示： 2 700-3 000 m和3 000 m以上海拔鼠形动物染蚤概率较2 400-2 700 m分别增加1.27和3.72倍;湿度高于70%时,鼠形动物染蚤概率较湿度≤70%时减少41%;与齐氏姬鼠的染蚤概率相比,中华姬鼠的染蚤概率降低50%,大绒鼠的染蚤概率增加79%;体长超过104 mm的鼠形动物染蚤概率较体长≤104 mm的鼠形动物染蚤概率增加76%;气温高于15℃时,鼠形动物染蚤数量较温度≤15℃时降低67%;成年鼠形动物的染蚤数量较未成年鼠形动物的染蚤数量增加2.25倍;与春季相比,夏季的染蚤数量增加2.00倍,秋季的染蚤概率减少48%,冬季的染蚤概率和染蚤数量分别增加1.44和1.06倍。【结论】玉龙鼠疫疫源地野外鼠形动物寄生蚤以特新蚤指名亚种、方叶栉眼蚤、无值大锥蚤、云南栉眼蚤为优势蚤种。鼠形动物寄生蚤丰盛度与海拔、季节、气温、湿度等环境气象因子及鼠形动物种类、体长、年龄等鼠形动物特征变量密切相关。     

The quantity of blood ingested by various species of fleas in an experiment

Data are given on the blood amount engorged by the fleas Xenopsylla conformis, X. cheopis, Leptopsylla segnis, Nosopsyllus laeviceps, N. consimilis, Ctenophthalmus golovi, Neopsylla setosa, Citellophilus tesquorum and Coptopsylla lamellifer. The average weight of blood portion in females of different species engorged for the first time ranged from 0.05 mg (X. conformis) to 0.72 mg (C. lamellifer). Females of most species, which had multiple blood meals, engorge more blood. Males engorge less blood than females and blood portions do not increase with age. In all sex-age groups most ectoparasites cease feeding spontaneously after having incomplete blood meal. Indices are suggested for the estimation of saturation rate in fleas during feeding.     

Bartonella and Rickettsia from fleas (Siphonaptera: Ceratophyllidae) of prairie dogs (Cynomys spp.) from the western United States

Fleas of prairie dogs have been implicated in the transmission of Bartonella spp. We used PCR to test DNA extracts from 47 fleas of prairie dogs from 6 states. We amplified DNA from 5 unique genotypes of Bartonella spp. and 1 Rickettsia sp. from 12 fleas collected in North Dakota, Oklahoma, Texas, and Wyoming. Sequences from the Bartonella spp. were similar, but not identical, to those from prairie dogs and their fleas in Colorado.     

Block formation capacity in fleas infected with a plague agent after their feeding with blood substitutes

By feeding plague infected Xenopsylla cheopis fleas with blood substitutes free of blood formed elements blocked fleas were obtained. These fleas are able to transmit plague agent to white mice and cause their mortality.     

云南居民区鼠类体外寄生蚤物种多样性调查

【目的】了解云南居民区鼠类寄生蚤的群落结构和空间分布特征。【方法】根据云南不同经纬度、 海拔等自然环境条件, 于2007年4月-2012年11月, 选取云南17个县(市) 居民区作为样区, 系统开展鼠类体外寄生蚤的调查, 运用群落结构指标对居民区鼠类寄生蚤的群落特征和沿环境梯度的空间分布进行研究。【结果】结果显示： 在调查的17个县(市)室内共检获鼠类体外寄生蚤521头, 隶属4科7亚科9属12种。居民区寄生蚤的纬度和垂直分布类似, 低纬度和低海拔范围的种类相对较少, 印鼠客蚤和缓慢细蚤是室内寄生蚤的优势种。相对纬度和海拔较高的区域, 居民区寄生蚤种类增多, 但优势种不突出; 在经度水平分布上, 蚤种类于99°~101°E经度带形成一个高峰, 室内寄生的优势种印鼠客蚤和缓慢细蚤几乎在所有经度带都可见到分布, 显示了较宽的生态幅。另外, 居民区寄生蚤物种丰富度和多样性指数水平分布（纬度）和垂直分布呈现为单峰格局, 总体显示随着纬度和海拔的升高, 先升高后降低的分布特征, 而在另一个水平分布（经度）, 则呈现由西向东呈递减的趋势。【结论】研究认为, 云南居民区蚤类的空间分布表现为独特的地理区域特征, 气候环境、 森林植被和人类生活生产方式通过影响蚤类栖息生境来影响蚤类的分布。     

Seasonal patterns of rodents, fleas and plague status in the Western Usambara Mountains, Tanzania

Field and commensal rodents were live-trapped at three villages in an active focus of plague (Yersinia pseudotuberculosis pestis) in Lushoto District, Western Usambara Mountains, Tanga Region, Tanzania, from December 1983 to November 1984. Their flea ectoparasites were collected, identified and counted. The rodent carcasses were serologically examined for specific plague antibodies and antigens, and bacteriologically examined for bipolar staining bacilli. A total of 1758 traps were set during the 12-month period and 924 animals were caught. From these, 1037 fleas were collected. Rattus rattus (L.), Praomys natalensis (Smith) and Lophuromys flavopunctatus Thomas comprised the largest proportions of the rodent population, while Dinopsyllus lypusus Jordan & Rothschild, Ctenophthalmus calceatus Waterston and Xenopsylla brasiliensis (Baker) were the dominant flea species. Rodents were most abundantly trapped during December and January. Flea indices were highest from December to May. Human plague was most active from November to March. Rodents contained plague antibodies every month except May and July, with a peak in September. Plague antigens and bipolar bacilli were detected in rodent organs during January-April. From the product of abundance and infection rate, the most prevalent rodent hosts of plague appeared to be R. rattus, Otomys angoniensis Wroughton, P. natalensis and Pelomys fallax (Peters). Continuous integrated control of rodents and fleas was recommended, reinforced by quarantine and maintenance of a surveillance service for clinical detection, diagnosis and treatment of patients in the plague endemic area.     

Detection of Rickettsia felis,Rickettsia typhi,Bartonella Species and Yersinia pestis in Fleas (Siphonaptera) from Africa

Little is known about the presence/absence and prevalence of Rickettsia spp, Bartonella spp. and Yersinia pestis in domestic and urban flea populations in tropical and subtropical African countries.

Methodology/Principal findings

Fleas collected in Benin, the United Republic of Tanzania and the Democratic Republic of the Congo were investigated for the presence and identity of Rickettsia spp., Bartonella spp. and Yersinia pestis using two qPCR systems or qPCR and standard PCR. In Xenopsylla cheopis fleas collected from Cotonou (Benin), Rickettsia typhi was detected in 1% (2/199), and an uncultured Bartonella sp. was detected in 34.7% (69/199). In the Lushoto district (United Republic of Tanzania), R. typhi DNA was detected in 10% (2/20) of Xenopsylla brasiliensis, and Rickettsia felis was detected in 65% (13/20) of Ctenocephalides felis strongylus, 71.4% (5/7) of Ctenocephalides canis and 25% (5/20) of Ctenophthalmus calceatus calceatus. In the Democratic Republic of the Congo, R. felis was detected in 56.5% (13/23) of Ct. f. felis from Kinshasa, in 26.3% (10/38) of Ct. f. felis and 9% (1/11) of Leptopsylla aethiopica aethiopica from Ituri district and in 19.2% (5/26) of Ct. f. strongylus and 4.7% (1/21) of Echidnophaga gallinacea. Bartonella sp. was also detected in 36.3% (4/11) of L. a. aethiopica. Finally, in Ituri, Y. pestis DNA was detected in 3.8% (1/26) of Ct. f. strongylus and 10% (3/30) of Pulex irritans from the villages of Wanyale and Zaa.

Conclusion

Most flea-borne infections are neglected diseases which should be monitored systematically in domestic rural and urban human populations to assess their epidemiological and clinical relevance. Finally, the presence of Y. pestis DNA in fleas captured in households was unexpected and raises a series of questions regarding the role of free fleas in the transmission of plague in rural Africa, especially in remote areas where the flea density in houses is high.     

Host location,survival and fecundity of the Oriental rat flea Xenopsylla cheopis (Siphonaptera: Pulicidae) in relation to black rat Rattus rattus (Rodentia: Muridae) host age and sex

Host choice and fecundity are two factors that may contribute to the variation in flea counts observed when assessing the potential risk of flea-borne transmission of pathogens from rodents to humans. Using the black rat, Rattus rattus Linnaeus, as host the effects of age and sex on host choice and fecundity of the Oriental rat flea, Xenopsylla cheopis Rothschild, were examined experimentally at 25 degrees C and 80% rh. During the first two days of emergence from cocoons, female fleas dominated the sex ratio by 4:1 but from the third day onwards this switched to a male-dominated sex ratio of 4:1. The sex of the flea did not influence their host-seeking behaviour. Newly emerged fleas of both sexes were not influenced by the rat's presence and at seven days old both sexes demonstrated similar levels of attraction toward the rat host. The sex of the rat did not affect flea host-seeking behaviour. There was a 50-70% decline in the initial number of adult fleas during the first week after their release onto a rat host, and this decline was greatest on juvenile rats. Flea fecundity was also significantly lower on juvenile rat hosts but no differences due to the sex of the rat were observed. This experimental study supports the hypothesis that differences in flea count due to host sex, reported in field surveys, result from sexual differences in host behaviour and not from discriminatory host-seeking behaviour by X. cheopis. Differences in flea count due to host age may be affected by differences in X. cheopis fecundity, which may itself be mediated by host behaviour such as grooming.     

HOST-SPECIFICITY AND HOST-SELECTION OF FLEAS IN FOCI OF HUMAN PLAGUE IN YUNNAN CHINA*

Abstract The host-specificity and the host-selection of 11 species of fleas collected from 47 species of small mammals in foci of human plague in Yunnan, China, were studied by using methods in the evaluation of ecological niche breadth and overlap. Levins' niche breadth was used for the host-specificity. while clip angle niche overlap and a fuzzy clustering analysis were used for host-selection. Of the 11 species of fleas, the host-specificity of Nosopsyllus elongatus puerensis and Xenopsylla cheopis are the highest (narrow niche breadth), and those of Aviostivalis klossi bispiniformis and Neopsylla stevensi sichuanyunnana the lowest (wide niche breadth). Of 11 species of fleas, the dominant host of X. cheopis (a very high effective vector of plague in the foci of human plague in Yunnan Province. China) is Rattw flavipectus (the main animal host and infectious source of plague in the foci). A high host-specificity of X. cheopis implies that X. cheopis mainly maintains or transmits the pathogen of plague among the individuals of its dominant species of host, R. flavipectus. The result of niche overlap analysis reveals that Ctenophthalrnus In-evipre jiciens and Ctenophthaltnus parcus have a similar host-selection while other species of fleas are quite different in their host selection.     

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats in the south of Brazil: a molecular study

Bartonella spp are the causative agent of cat scratch disease in humans. Cats are the natural reservoir of these bacteria and may infect humans through scratches, bites or fleas. Blood samples from 47 cats aged up to 12 months were collected for this study. All animals were lodged in municipal animal shelters in the Vale do Sinos region, Rio Grande do Sul, Brazil. Bartonella spp were detected by genus-specific polymerase chain reaction (PCR) and when the PCR was positive, the species were determined by DNA sequencing. A Giemsa-stained blood smear was also examined for the presence of intraerythrocytic elements suggestive of Bartonella spp infection. Phylogenetic analysis was also performed for all positive samples. Using molecular detection methods, Bartonella spp were detected in 17.02% (8/47) of the samples. In seven out of eight samples confirmed to be positive for Bartonella spp, blood smear examination revealed the presence of intraerythrocytic elements suggestive of Bartonella spp. Phylogenetic analysis characterized positive samples as Bartonella henselae (5) or Bartonella clarridgeiae (3). To the best of our knowledge, this is the first molecular study demonstrating the presence of Bartonella spp in cats from the Southern Region of Brazil.     

Pathogenic action of the plague microbe on the flea Xenopsylla cheopis and the ultrastructure of the causative agent at various times of its stay in the vector

Pathology of gastro-intestinal tract of Xenopsylla cheopis fleas infected with plague microbe was determined by means of electron microscopy. Ultrastructure of plague microbe during different periods of its stay in the vector was studied.     

Epidemiological prediction of the distribution of insects of medical significance: comparative distributions of fleas and sucking lice on the rat host Rattus norvegicus in Yunnan Province, China

Determining the distribution patterns of ectoparasites is important for predicting the spread of vector-borne diseases. A simple epidemiological model was used to compare the distributions of two different taxa of ectoparasitic insects, sucking lice (Insecta: Siphonaptera) and fleas (Insecta: Anoplura), on the same rodent host, Rattus norvegicus Berkenhout (Rodentia: Muridae), in Yunnan Province, China. Correlations between mean abundance and prevalence were determined. Both fleas and sucking lice were aggregated on their hosts, and sucking lice showed a higher degree of aggregation than fleas. The prevalence of both fleas and sucking lice increased with log-transformed mean abundance and a highly linear correlation and modelling efficiency of predicted prevalence against observed prevalence were obtained. The results demonstrate that prevalence can be explained simply by mean abundance.     

Sex ratio in flea infrapopulations: number of fleas, host gender and host age do not have an effect

This study set out to determine whether the sex ratio of fleas collected from host bodies is a reliable indicator of sex ratio in the entire flea population. To answer this question, previously published data on 18 flea species was used and it was tested to see whether a correlation exists between the sex ratio of fleas collected from host bodies and the sex ratio of fleas collected from host burrows. Across species, the female:male ratio of fleas on hosts correlated strongly with the female:male ratio of fleas in their burrows, with the slope of the regression overlapping 1. Controlling for flea phylogeny by independent contrasts produced similar results. It was also ascertained whether a host individual is a proportional random sampler of male and female fleas and whether the sex ratio in flea infrapopulations depends on the size of infrapopulations and on the gender and age of a host. Using field data, the sex ratio in infrapopulations of 7 flea species parasitic on 4 rodent species was analysed. Populations of 3 species (Nosopsyllus iranus, Parapulex chephrenis and Xenopsylla conformis) were significantly female-biased, whereas male bias was found in 1 species (Synosternus cleopatrae). In general, the sex ratio of fleas collected from an individual rodent did not differ significantly from the sex ratio in the entire flea population. Neither host gender, and age nor number of fleas co-occurring on a host affected (a) the sex ratio in flea infrapopulations and (b) the probability of an infrapopulation to be either female- or male-biased.     

Characteristics of the multiplication of a virulent strain of the plague microbe in Xenopsylla cheopis fleas infected parenterally

Xenopsylla cheopis fleas infected parenterally with the virulent strain of plague microbe of gerbil variant preserved the agent to the end of their lives. In the body cavity the microbes retained their ability for reproduction which was, however, limited. During the first seven days after the infection the number of microbes slightly increased and later became stabilized. Its mean indices (mean g) varied within the limits of 500 to 2000 microbe cells per 1 individual, maximum index rarely exceeded 30 000 microbe cells. Parenteral infection with plague agent did not affect essentially the longevity of fleas.     

Notes on fleas (Siphonaptera) in plague foci on the Tay Nguyen plateau (Vietnam)]

The fleas of wild and commensal small mammals, domestic animals (dogs, cats) and free-living flea forms in houses have been collected in plague nidi of Tay Nguyen plate, Dak-Lak province, Vietnam. Pulex irritans, Ctenocephalides felis felis, Ct. felis orientis were found in the houses on dogs, cats and on the ground floor. Commensal rats in populated areas were infested by Xenopsylla cheopis and rarely by Lentistivalius klossi. The agricultural zone was inhabited by both home and wild animals such as commensal, savannah and forest-dwelling small mammals. The flea fauna of this zone is presented by X. cheopis and L. klossi. In the tropical forest surrounding villages four of the flea species were found: X. vexabilis, a specific parasite of the forest-dwelling rat Berylmys berdmorei, L. klossi found on six species of forest small mammals, Acropsylla girshami from Berylmys bowersii and Pariodontis subjugis from Hystrix brachyura. The agricultural zone is the most possible place of commensal and forest-dwelling small mammals contact, where the latter can get plague microbe.