Genetic diversity of Bartonella species in small mammals in the Qaidam Basin, western China.

Investigation of the prevalence and diversity of Bartonella infections in small mammals in the Qaidam Basin, western China, could provide a scientific basis for the control and prevention of Bartonella infections in humans. Accordingly, in this study, small mammals were captured using snap traps in Wulan County and Ge'ermu City, Qaidam Basin, China. Spleen and brain tissues were collected and cultured to isolate Bartonella strains. The suspected positive colonies were detected with polymerase chain reaction amplification and sequencing of gltA, ftsZ, RNA polymerase beta subunit (rpoB) and ribC genes. Among 101 small mammals, 39 were positive for Bartonella, with the infection rate of 38.61%. The infection rate in different tissues (spleens and brains) (χ2 = 0.112, P = 0.738) and gender (χ2 = 1.927, P = 0.165) of small mammals did not have statistical difference, but that in different habitats had statistical difference (χ2 = 10.361, P = 0.016). Through genetic evolution analysis, 40 Bartonella strains were identified (two different Bartonella species were detected in one small mammal), including B. grahamii (30), B. jaculi (3), B. krasnovii (3) and Candidatus B. gerbillinarum (4), which showed rodent-specific characteristics. B. grahamii was the dominant epidemic strain (accounted for 75.0%). Furthermore, phylogenetic analysis showed that B. grahamii in the Qaidam Basin, might be close to the strains isolated from Japan and China. Overall, we observed a high prevalence of Bartonella infection in small mammals in the Qaidam Basin. B. grahamii may cause human disease, and the pathogenicity of the others Bartonella species needs further study, the corresponding prevention and control measures should be taken into consideration.

Molecular detection and clinical characteristics of Bartonella bacilliformis, Leptospira spp., and Rickettsia spp. in the Southeastern Peruvian Amazon basin.

BACKGROUND: Acute febrile illness (AFI) represent a significant health challenge in the Peruvian Amazon basin population due to their diverse etiologies and the unavailability of specific on-site diagnostic methods, resulting in underreporting of cases. In Peru, one of the most endemic regions to dengue and leptospirosis is Madre de Dios, a region also endemic to emergent bacterial etiologic agents of AFI, such as bartonellosis and rickettsiosis, whose prevalence is usually underreported. We aimed to molecularly identify the presence of Leptospira spp., Bartonella bacilliformis, and Rickettsia spp. by Polymerase Chain Reaction in serum samples from patients with AFI from Puerto Maldonado-Madre de Dios in Peru. METHODS: Serum samples from patients with acute febrile illness were analyzed by real-time PCR for detecting the presence of Bartonella bacilliformis, Leptospira spp. and Rickettsia spp. RESULTS: Bartonella bacilliformis was the most prevalent bacteria identified in 21.6% (30/139) of the samples, followed by Leptospira spp. in 11.5% (16/139) and Rickettsia spp. in 6.5% (9/139) of the samples. No co-infections were observed between these bacteria. The most frequent symptoms associated with fever among all groups, were headaches, myalgias, and arthralgias. We found no statistically significant differences in the clinical presentation between patients infected with each bacterium. CONCLUSIONS: In a previous study, we shown the presence of dengue, chikungunya, Zika and oropouche virus. We were able to identify these pathogens in 29.5% of all the samples, with chikungunya and OROV as the most frequently found in 9.4 and 8.6% of all the samples, respectively. In this study we show that B. bacilliformis (21.6%), Leptospira spp. (11.5%) and Rickettsia spp. (6.5%) accounted for the main etiologies of AFI in samples from Puerto Maldonado-Madre de Dios, Perú. Our analysis of their clinical presentation, further shows the importance of implementing more sensitive and specific on-site diagnostic tools in the national surveillance programs.This study confirms that the un-specificity of signs and symptoms is not only associated with arboviral infections, but also with the clinical presentation of endemic bacterial infections.

Bartonella Infection in Hematophagous, Insectivorous, and Phytophagous Bat Populations of Central Mexico and the Yucatan Peninsula.

Although emerging nonviral pathogens remain relatively understudied in bat populations, there is an increasing focus on identifying bat-associated bartonellae around the world. Many novel Bartonella strains have been described from both bats and their arthropod ectoparasites, including Bartonella mayotimonensis, a zoonotic agent of human endocarditis. This cross-sectional study was designed to describe novel Bartonella strains isolated from bats sampled in Mexico and evaluate factors potentially associated with infection. A total of 238 bats belonging to seven genera were captured in five states of Central Mexico and the Yucatan Peninsula. Animals were screened by bacterial culture from whole blood and/or polymerase chain reaction of DNA extracted from heart tissue or blood. Bartonella spp. were isolated or detected in 54 (22.7%) bats, consisting of 41 (38%) hematophagous, 10 (16.4%) insectivorous, and three (4.3%) phytophagous individuals. This study also identified Balantiopteryx plicata as another possible bat reservoir of Bartonella. Univariate and multivariate logistic regression models suggested that Bartonella infection was positively associated with blood-feeding diet and ectoparasite burden. Phylogenetic analysis identified a number of genetic variants across hematophagous, phytophagous, and insectivorous bats that are unique from described bat-borne Bartonella species. However, these strains were closely related to those bartonellae previously identified in bat species from Latin America.

Evolutionary Dynamics of Pathoadaptation Revealed by Three Independent Acquisitions of the VirB/D4 Type IV Secretion System in Bartonella.

The α-proteobacterial genus Bartonella comprises a group of ubiquitous mammalian pathogens that are studied as a model for the evolution of bacterial pathogenesis. Vast abundance of two particular phylogenetic lineages of Bartonella had been linked to enhanced host adaptability enabled by lineage-specific acquisition of a VirB/D4 type IV secretion system (T4SS) and parallel evolution of complex effector repertoires. However, the limited availability of genome sequences from one of those lineages as well as other, remote branches of Bartonella has so far hampered comprehensive understanding of how the VirB/D4 T4SS and its effectors called Beps have shaped Bartonella evolution. Here, we report the discovery of a third repertoire of Beps associated with the VirB/D4 T4SS of B. ancashensis, a novel human pathogen that lacks any signs of host adaptability and is only distantly related to the two species-rich lineages encoding a VirB/D4 T4SS. Furthermore, sequencing of ten new Bartonella isolates from under-sampled lineages enabled combined in silico analyses and wet lab experiments that suggest several parallel layers of functional diversification during evolution of the three Bep repertoires from a single ancestral effector. Our analyses show that the Beps of B. ancashensis share many features with the two other repertoires, but may represent a more ancestral state that has not yet unleashed the adaptive potential of such an effector set. We anticipate that the effectors of B. ancashensis will enable future studies to dissect the evolutionary history of Bartonella effectors and help unraveling the evolutionary forces underlying bacterial host adaptation.

Novel Bartonella Species in Insectivorous Bats, Northern China.

Bartonella species are emerging human pathogens. Bats are known to carry diverse Bartonella species, some of which are capable of infecting humans. However, as the second largest mammalian group by a number of species, the role of bats as the reservoirs of Bartonella species is not fully explored, in term of their species diversity and worldwide distribution. China, especially Northern China, harbors a number of endemic insectivorous bat species; however, to our knowledge, there are not yet studies about Bartonella in bats in China. The aim of the study was to investigate the prevalence and genetic diversity of Bartonella species in bats in Northern China. Bartonella species were detected by PCR amplification of gltA gene in 25.2% (27/107) bats in Mengyin County, Shandong Province of China, including 1/3 Rhinolophus ferrumequinum, 2/10 Rhinolophus pusillus, 9/16 Myotis fimbriatus, 1/5 Myotis ricketti, 14/58 Myotis pequinius. Phylogenetic analysis showed that Bartonella species detected in bats in this study clustered into ten groups, and some might be novel Bartonella species. An association between Bartonella species and bat species was demonstrated and co-infection with different Bartonella species in a single bat was also observed. Our findings expanded our knowledge on the genetic diversity of Bartonella in bats, and shed light on the ecology of bat-borne Bartonella species.

Prevalence and genetic diversity of Bartonella strains in rodents from northwestern Mexico.

Bartonella infections were investigated in wild rodents from northwestern Chihuahua, Mexico. A total of 489 rodents belonging to 14 species were surveyed in four areas. Bartonella bacteria were cultured from 50.1% of rodent samples (245/489). Infection rates ranged from 0% to 83.3% per rodent species, with no significant difference between sites except for Cynomys ludovicianus. Phylogenetic analyses of the citrate synthase gene (gltA) of the Bartonella isolates revealed 23 genetic variants (15 novel and 8 previously described), clustering into five phylogroups. Three phylogroups were associated with Bartonella vinsonii subsp. vinsonii, B. vinsonii subsp. arupensis, and B. washoensis, respectively. The other two phylogroups were not genetically related to any known Bartonella species. The genetic variants and phylogenetic groups exhibited a high degree of host specificity, mainly at the genus and family levels. This is the first study that describes the genetic diversity of Bartonella strains in wild rodents from Mexico. Considering that some variants found in this study are associated with Bartonella species that have been reported as zoonotic, more investigations are needed to further understand the ecology of Bartonella species in Mexican wildlife and their implications for human health.

Accurate prediction of bacterial type IV secreted effectors using amino acid composition and PSSM profiles.

MOTIVATION: Various human pathogens secret effector proteins into hosts cells via the type IV secretion system (T4SS). These proteins play important roles in the interaction between bacteria and hosts. Computational methods for T4SS effector prediction have been developed for screening experimental targets in several isolated bacterial species; however, widely applicable prediction approaches are still unavailable RESULTS: In this work, four types of distinctive features, namely, amino acid composition, dipeptide composition, .position-specific scoring matrix composition and auto covariance transformation of position-specific scoring matrix, were calculated from primary sequences. A classifier, T4EffPred, was developed using the support vector machine with these features and their different combinations for effector prediction. Various theoretical tests were performed in a newly established dataset, and the results were measured with four indexes. We demonstrated that T4EffPred can discriminate IVA and IVB effectors in benchmark datasets with positive rates of 76.7% and 89.7%, respectively. The overall accuracy of 95.9% shows that the present method is accurate for distinguishing the T4SS effector in unidentified sequences. A classifier ensemble was designed to synthesize all single classifiers. Notable performance improvement was observed using this ensemble system in benchmark tests. To demonstrate the model's application, a genome-scale prediction of effectors was performed in Bartonella henselae, an important zoonotic pathogen. A number of putative candidates were distinguished. AVAILABILITY: A web server implementing the prediction method and the source code are both available at http://bioinfo.tmmu.edu.cn/T4EffPred.

Deciphering bartonella diversity, recombination, and host specificity in a rodent community.

Host-specificity is an intrinsic feature of many bacterial pathogens, resulting from a long history of co-adaptation between bacteria and their hosts. Alpha-proteobacteria belonging to the genus Bartonella infect the erythrocytes of a wide range of mammal orders, including rodents. In this study, we performed genetic analysis of Bartonella colonizing a rodent community dominated by bank voles (Myodes glareolus) and wood mice (Apodemus sylvaticus) in a French suburban forest to evaluate their diversity, their capacity to recombine and their level of host specificity. Following the analysis of 550 rodents, we detected 63 distinct genotypes related to B. taylorii, B. grahamii, B. doshiae and a new B. rochalimae-like species. Investigating the most highly represented species, we showed that B. taylorii strain diversity was markedly higher than that of B. grahamii, suggesting a possible severe bottleneck for the latter species. The majority of recovered genotypes presented a strong association with either bank voles or wood mice, with the exception of three B. taylorii genotypes which had a broader host range. Despite the physical barriers created by host specificity, we observed lateral gene transfer between Bartonella genotypes associated with wood mice and Bartonella adapted to bank voles, suggesting that those genotypes might co-habit during their life cycle.

Identification of Bartonella Trw host-specific receptor on erythrocytes.

Each Bartonella species appears to be highly adapted to one or a limited number of reservoir hosts, in which it establishes long-lasting intraerythrocytic bacteremia as the hallmark of infection. Recently, we identified Trw as the bacterial system involved in recognition of erythrocytes according to their animal origin. The T4SS Trw is characterized by a multiprotein complex that spans the inner and outer bacterial membranes, and possesses a hypothetical pilus structure. TrwJ, I, H and trwL are present in variable copy numbers in different species and the multiple copies of trwL and trwJ in the Bartonella trw locus are considered to encode variant forms of surface-exposed pilus components. We therefore aimed to identify which of the candidate Trw pilus components were located on the bacterial surface and involved in adhesion to erythrocytes, together with their erythrocytic receptor. Using different technologies (electron microscopy, phage display, invasion inhibition assay, far western blot), we found that only TrwJ1 and TrwJ2 were expressed and localized at the cell surface of B. birtlesii and had the ability to bind to mouse erythrocytes, and that their receptor was band3, one of the major outer-membrane glycoproteins of erythrocytes, (anion exchanger). According to these results, we propose that the interaction between TrwJ1, TrwJ2 and band 3 leads to the critical host-specific adherence of Bartonella to its host cells, erythrocytes.

Bartonella species in dogs and their ectoparasites from Khon Kaen Province, Thailand.

In order to access the prevalence of Bartonella species in dogs, whole blood and any associated ectoparasites were collected from 164 dogs with owners in 25 villages throughout Khon Kaen Province. DNA was extracted from dog blood, 92 ticks (Rhipicephalus sanguineus) and 137 fleas (Ctenocephalides spp) and screened by PCR using intergenic spacer region and citrate synthase gene primers. B. clarridgeiae DNA was detected in blood of 3 dogs, 4 C. felis and 1 C. canis; B. rochalimae DNA was found in 1 tick; and B. vinsonii subsp vinsonii DNA was found in 2 C. felis. The findings indicate that dogs residing in northeast Thailand are exposed to diverse Bartonella species that are also potential human pathogens.

Genetic characterization of flea-derived Bartonella species from native animals in Australia suggests host-parasite co-evolution.

Fleas are important arthropod vectors for a variety of diseases in veterinary and human medicine, and bacteria belonging to the genus Bartonella are among the organisms most commonly transmitted by these ectoparasites. Recently, a number of novel Bartonella species and novel species candidates have been reported in marsupial fleas in Australia. In the present study the genetic diversity of marsupial fleas was investigated; 10 species of fleas were collected from seven different marsupial and placental mammal hosts in Western Australia including woylies (Bettongia penicillata), western barred bandicoots (Perameles bougainville), mardos (Antechinus flavipes), bush rats (Rattus fuscipes), red foxes (Vulpes vulpes), feral cats (Felis catus) and rabbits (Oryctolagus cuniculus). PCR and sequence analysis of the cytochrome oxidase subunit I (COI) and the 18S rRNA genes from these fleas was performed. Concatenated phylogenetic analysis of the COI and 18S rRNA genes revealed a close genetic relationship between marsupial fleas, with Pygiopsylla hilli from woylies, Pygiopsylla tunneyi from western barred bandicoots and Acanthopsylla jordani from mardos, forming a separate cluster from fleas collected from the placental mammals in the same geographical area. The clustering of Bartonella species with their marsupial flea hosts suggests co-evolution of marsupial hosts, marsupial fleas and Bartonella species in Australia.

Enfoque genético evolutivo de la enfermedad de Carrión: polimorfismos de los grupos sanguíneos MN, Ss, Diego, Duffy y fases clínicas en zonas endémicas de Bagua, Amazonas / Evolutionary genetic approach of Carrion's disease: MN, Ss, Diego, Duffy blood groups polymorphisms and clinical stages in endemic areas of Bagua, Amazonas, Peru

Objetivos: Establecer la relación entre las frecuencias fenotípicas y/o alélicas de los grupos sanguíneos MN, Ss, Diego, Duffy y las fases clínicas de la enfermedad de Carrión, interpretado en un contexto genético coevolutivo del hospedero amerindio con la bacteria Bartonella bacilliformis. Diseño: Estudio asociativo y analítico. Lugar: Instituto de Medicina Tropical Daniel A. Carrión, Facultad de Medicina, Universidad Nacional Mayor de San Marcos, Lima, Perú. Participantes: Pobladores de Bagua Grande, Amazonas, Perú. Intervenciones: Se determinó los grupos sanguíneos MN, Ss, Diego y Duffy, según metodología estándar, en 40 pobladores de las zonas de Tomocho-Collicate-Vista Hermosa (antecedente de casos en fase verrucosa o benigna, sin aparente fase aguda previa) y 36 pobladores de la zona de Miraflores (antecedente de casos solo en fase aguda), de Bagua Grande, Amazonas. Principales medidas de resultados: Frecuencias fenotípicas y alélicas de los referidos grupos sanguíneos en las localidades estudiadas. Resultados: No existieron diferencias significativas entre las frecuencias fenotípicas, genotípicas y/o alélicas de los grupos sanguíneos MN, Ss, Diego, Duffy en las zonas de Tomocho-Collicate-Vista Hermosa (predominancia de fase verrucosa) y en la zona de Miraflores (predominancia de fase aguda). La más variable fue la del grupo MN, pero encontrándose en equilibrio de Hardy-Weinberg. Conclusiones: Los marcadores sanguíneos Ss, Diego y Duffy -inclusive el MN, que aparentemente mostró la mayor variabilidad-, no tuvieron relación con las fases clínicas de la enfermedad de Carrión, en las localidades estudiadas. Sin embargo, en el contexto de un enfoque genético evolutivo, es necesario evaluar dicha asociación en otras zonas endémicas del país, así como la susceptibilidad, resistencia y otras carácterísticas clínicas de esta ancestral enfermedad.

Objectives: To determine the relationship between MN, Ss, Diego, Duffy blood groups phenotypic or alleles frequencies and clinical phases of Carrion's disease, interpreted in a coevolutive genetic context of amerindian hosts with Bartonella bacilliformis. Design: Associative and analytical study. Setting: Daniel A. Carrion Tropical Medicine Institute, Faculty of Medicine, Universidad Nacional Mayor de San Marcos, Lima, Peru. Participants: Bagua Grande, Amazonas, Peru settlers. Interventions: MN, Ss, Duffy and Diego blood groups were determined using standard methodology in 40 Tomocho-Collicate-Vista Hermosa settlers (background of benign or verrucose phase cases without apparent acute phase) and 36 residents of Miraflores area (background of only acute phase cases), Bagua Grande, Amazonas. Main outcomes measures: Phenotypic and allele frequencies of these blood groups in the localities studied. Results: There were no significant differences between the phenotypic, genotypic or alleles frequencies of the MN, Ss, Diego, Duffy blood groups in the Tomocho-Collicate-Vista Hermosa areas (predominance of the verrucose phase) and the Miraflores area (predominance of the acute phase), Bagua Grande, Amazonas. The most variable frequency was found in the MN group, but it was within Hardy-Weinberg equilibrium. Conclusions: Ss, Duffy and Diego blood markers, including the most variable MN, were unrelated to Carrion's disease clinical phases in the localities studied. However, in the context of evolutionary genetic approach, association must be assessed in other endemic areas of the country, as well as susceptibility, resistance and other clinical features of this ancient disease.

Multi-locus sequence analysis reveals host specific association between Bartonella washoensis and squirrels.

To clarify phylogenetic relationships and genetic diversity among Bartonella washoensis strains obtained from squirrels, multi-locus sequence analysis (MLSA) with the 16S rRNA, ftsZ, gltA, groEL, ribC, and rpoB genes was applied for 20 strains of B. washoensis isolated from five genera of squirrels (Tamias, Tamiasciurus, Glaucomys, Sciurus, and Spermophilus) within the family Sciuridae. Sequence similarities in the concatenated sequences of B. washoensis strains from squirrels of different genera ranged from 94.7% (Sciurus vs. Spermophilus) to 98.4% (Tamiasciurus vs. Glaucomys). Phylogenetic trees based on the concatenated sequences revealed that B. washoensis strains formed five distinct clades and each clade correlated with the genus of squirrel from which the strains were originally obtained. The discrimination was supported by 100% bootstrap values and posterior probabilities, respectively. These results suggest that B. washoensis strains may have co-speciated with their squirrel hosts and provide new insights into the application of the MLSA to identify sources of B. washoensis infection with accuracy.

Epidemiology of Bartonella infection in rodents and shrews in Taiwan.

During the period of August 2002 and November 2004, an epidemiological investigation for Bartonella infection was conducted in small mammals in Taiwan. Using whole blood culture on chocolate agar plates, Bartonella species were successfully isolated from 41.3% of the 310 animals tested. The isolation rate of Bartonella species varied among different animal species, including 52.7% of the 169 Rattus norvegicus, 28.6% of the 126 Sucus murinus, 10% of the 10 Rattus rattus and 66.7% of the three Rattus losea. Bacteremia prevalence also varied with the origin of the animals, as 56.2% of the animals captured on farms, 38.6% of the ones captured at harbour sites and 11.8% of the animals captured from urban areas were bacteremic. Through molecular analysis of the gltA gene and 16S/23S intergenic spacer region, genetic diversity of Bartonella organisms was identified, including strains closely related to Bartonella tribocorum, Bartonella grahamii, Bartonella elizabethae, Bartonella phoceensis and Bartonella rattimassiliensis. Moreover, this is the first report of zoonotic B. elizabethae and B. grahamii identified in R. losea, the lesser rice-field rat. Various Bartonella species were identified in R. norvegicus, compared to 97.2% of Suncus murinus with unique Bartonella species. By indirect immunofluorescence antibody test, using various rodent Bartonella species as antigens, consistently low percentage of seropositivity implied that small mammals may play a role as competent reservoirs of Bartonella species in Taiwan. Future studies need to be conducted to determine whether these Bartonella species would be responsible for human cases of unknown fever or febrile illness in Taiwan, especially zoonotic B. elizabethae and B. grahamii.

Detection of Bartonella spp. in wild rodents in Israel using HRM real-time PCR.

The prevalence of Bartonella spp. in wild rodents was studied in 19 geographical locations in Israel. One hundred and twelve rodents belonging to five species (Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus) were included in the survey. In addition, 156 ectoparasites were collected from the rodents. Spleen sample from each rodent and the ectoparasites were examined for the presence of Bartonella DNA using high resolution melt (HRM) real-time PCR. The method was designed for the simultaneous detection and differentiation of eight Bartonella spp. according to the nucleotide variation in each of two gene fragments (rpoB and gltA) and the 16S-23S intergenic spacer (ITS) locus, using the same PCR protocol which allowed the simultaneous amplification of the three different loci. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). In addition, 15 of 34 (44%) flea pools harbored Bartonella DNA. Only rat flea (Xenopsyla cheopis) pools collected from black rats (R. rattus) were positive for Bartonella DNA. The Bartonella sp. detected in spleen samples from black rats (R. rattus) was closely related to both B. tribocorum and B. elizabethae. The species detected in the Cairo spiny mouse (A. cahirinus) spleen sample was closely related to the zoonotic pathogen, B. elizabethae. These results indicate that Bartonella species are highly prevalent in suburban rodent populations and their ectoparasites in Israel. Further investigation of the prevalence and zoonotic potential of the Bartonella species detected in the black rats and the Cairo spiny mouse is warranted.

Emerging Bartonella in humans and animals in Asia and Australia.

Bartonella species, belonging to the alpha 2 subgroup of Proteobacteria, have either been considered or established as potential human and mammal pathogens. Five novel species of Bartonella have been reported in Thailand and Australia. Recently, three strains of B. tamiae were isolated from febrile illness patients in Thailand, while B. australis was isolated from kangaroos, and B. coopersplainsensis, B. queenslandensis, and B. rattiaustraliensis were isolated from rats in Australia. The 17 novel Bartonella strains isolated from rodents in southern China that were identified using the partial citrate synthase gene (gltA) sequence displayed a similar genetic diversity, as compared to those obtained from rodents captured in northern Thailand. Herein, the authors review and discuss the few available reports on Bartonella infection in order to raise awareness of Bartonella infection transmitted from mammalian reservoirs to humans via arthropod ectoparasitic vectors such as fleas, ticks, and lice in Asia and Australia. The identification of Bartonella species on these continents was reported in eastern Asia (China, Japan, Korea, Russia, and Taiwan), south central Asia (Afghanistan, Bangladesh, India, and Nepal), southeast Asia (Indonesia, Philippines, Singapore, and Thailand), the Middle East (Israel and Jordan), and Australia. The rate of Bartonella infection was found to be high in arthropod ectoparasitic vectors, mammals, and febrile patients in these tropical zones.

Major histocompatibility complex (MHC) heterozygote superiority to natural multi-parasite infections in the water vole (Arvicola terrestris).

The fundamental role of the major histocompatibility complex (MHC) in immune recognition has led to a general consensus that the characteristically high levels of functional polymorphism at MHC genes is maintained by balancing selection operating through host-parasite coevolution. However, the actual mechanism by which selection operates is unclear. Two hypotheses have been proposed: overdominance (or heterozygote superiority) and negative frequency-dependent selection. Evidence for these hypotheses was evaluated by examining MHC-parasite relationships in an island population of water voles (Arvicola terrestris). Generalized linear mixed models were used to examine whether individual variation at an MHC class II DRB locus explained variation in the individual burdens of five different parasites. MHC genotype explained a significant amount of variation in the burden of gamasid mites, fleas (Megabothris walkeri) and nymphs of sheep ticks (Ixodes ricinus). Additionally, MHC heterozygotes were simultaneously co-infected by fewer parasite types than homozygotes. In each case where an MHC-dependent effect on parasite burden was resolved, the heterozygote genotype was associated with fewer parasites, and the heterozygote outperformed each homozygote in two of three cases, suggesting an overall superiority against parasitism for MHC heterozygote genotypes. This is the first demonstration of MHC heterozygote superiority against multiple parasites in a natural population, a mechanism that could help maintain high levels of functional MHC genetic diversity in natural populations.

Bartonella species in blood of immunocompetent persons with animal and arthropod contact.

Using PCR in conjunction with pre-enrichment culture, we detected Bartonella henselae and B. vinsonii subspecies berkhoffii in the blood of 14 immunocompetent persons who had frequent animal contact and arthropod exposure.

A Bartonella vinsonii berkhoffii typing scheme based upon 16S-23S ITS and Pap31 sequences from dog, coyote, gray fox, and human isolates.

Since the isolation of Bartonella vinsonii subspecies berkhoffii from a dog with endocarditis in 1993, this organism has emerged as an important pathogen in dogs and as an emerging pathogen in people. Current evidence indicates that coyotes, dogs and gray foxes potentially serve as reservoir hosts. Based upon sequence differences within the 16S-23S ITS region and Pap31 gene, we propose a classification scheme that divides B. vinsonii subsp. berkhoffii isolates into four distinct types. Two conserved sequences, of 37 and 18 bp, respectively, are differentially present within the ITS region of each of the four B. vinsonii subsp. berkhoffii types. To date, B. vinsonii berkhoffii types I, II, and III have been identified in the US, type III in Europe and type IV in Canada. Based upon the proposed genotyping scheme, the geographic distribution of B. vinsonii berkhoffii types needs to be more thoroughly delineated in future molecular epidemiological studies involving Bartonella infection in coyotes, dogs, gray foxes, human beings and potentially other animals or in arthropod vectors. Strain typing may help to better define the reservoir potential, carriership patterns, modes of transmission, and geographic distribution for each B. vinsonii berkhoffii type.