Environmental and sociodemographic factors associated with zoonotic pathogen occurrence in Norway rats (Rattus norvegicus) from Windsor, Ontario.

AIMS: Rat-associated zoonotic pathogen transmission at the human-wildlife interface is a public health concern in urban environments where Norway rats (Rattus norvegicus) thrive on abundant anthropogenic resources and live in close contact with humans and other animal species. To identify potential factors influencing zoonotic pathogen occurrence in rats, we investigated associations between environmental and sociodemographic factors and Leptospira interrogans and Bartonella spp. infections in rats from Windsor, Ontario, Canada, while controlling for the potential confounding effects of animal characteristics (i.e., sexual maturity and body condition). METHODS AND RESULTS: Between November 2018 and June 2021, 252 rats were submitted by collaborating pest control professionals. Kidney and spleen samples were collected for L. interrogans and Bartonella spp. PCR and sequencing, respectively. Of the rats tested by PCR, 12.7% (32/252) were positive for L. interrogans and 16.3% (37/227) were positive for Bartonella species. Associations between infection status and environmental and sociodemographic variables of interest were assessed via mixed multivariable logistic regression models with a random intercept for social group and fixed effects to control for sexual maturity and body condition in each model. The odds of L. interrogans infection were significantly higher in rats from areas with high building density (odds ratio [OR]: 3.76; 95% CI: 1.31-10.79; p = 0.014), high human population density (OR: 3.31; 95% CI: 1.20-9.11; p = 0.021), high proportion of buildings built in 1960 or before (OR: 11.21; 95% CI: 2.06-60.89; p = 0.005), and a moderate number of reports of uncollected garbage compared to a low number of reports (OR: 4.88; 95% CI: 1.01-23.63; p = 0.049). A negative association was observed between median household income and Bartonella spp. infection in rats (OR: 0.26; 95% CI: 0.08-0.89; p = 0.031). CONCLUSIONS: Due to the complexity of the ecology of rat-associated zoonoses, consideration of environmental and sociodemographic factors is of critical importance to better understand the nuances of host-pathogen systems and inform how urban rat surveillance and intervention efforts should be distributed within cities.

Assessment of tick-borne pathogens presence in Dermacentor reticulatus ticks in north-eastern Poland.

PURPOSE: Dermacentor reticulatus is the second most common tick species in Poland after Ixodes ricinus. The aim of the study was to analyze the presence of pathogen DNA in D. reticulatus. MATERIALS AND METHODS: Ticks were collected in The Protected Landscape Area of the Bug and Nurzec Valley (52°40' N and 22°28' E) between 2016 and 2017. End-point PCR for Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia spp., Rickettsia spp., Bartonella spp. and Coxiella burnetii detection was performed. RESULTS: Tick-borne pathogens' DNA was detected in 11.3% of 301 ticks: B. burgdorferi s.l. in 3.6%, Babesia spp. in 6.3%, A. phagocytophilum in 0.7% and B. burgdorferi s.l.-Babesia spp. co-infection in 0.7%. In all 21 Babesia spp. positive samples, sequence analysis confirmed the presence of Babesia canis with an 80.3%-98.3% homology with the B. canis sequences in GenBank. C. burnetii, Bartonella spp., and Rickettsia spp. DNA were not detected. CONCLUSIONS: Dermacentor reticulatus from north-eastern Poland were found to carry three of the most common tick-borne pathogens (B. burgdorferi s.l., Babesia canis, A. phagocytophilum) which lead to single and mixed infections. Babesia canis was the most prevalent pathogen identified in D. reticulatus.

Molecular assessment of Bartonella in Gerbillus nanus from Saudi Arabia reveals high levels of prevalence, diversity and co-infection.

Bartonellae bacteria are associated with several re-emerging human diseases. These vector-borne pathogens have a global distribution, yet data on Bartonella prevalence and diversity in the Arabian Peninsula are limited. In this study we assessed the Bartonella infection status of the Baluchistan gerbil (Gerbillus nanus), a species associated with pastoral communities throughout the Middle East region, using a multi-gene PCR screening approach. The results demonstrated that 94 (68.1%) of the 138 gerbils trapped on a monthly basis, over a period of one year, were PCR-positive. Sequencing of the gltA gene region confirmed the presence of four discrete Bartonella lineages (I-IV) and high levels of co-infection (33.0%). Each of the four lineages, varied in overall abundance (7.5%-47.9%) and had discernible seasonal peaks. Bartonella status was significantly correlated with ectoparasite presence, but not with sex, nor with season. Statistical analyses further revealed that co-infected individuals had a significantly higher relative body condition. Multi-locus sequence analysis (MLSA) performed with a concatenated dataset of three genetic loci (gltA, nuoG, and rpoB), 1452 nucleotides (nt) in length confirmed that lineage IV, which occurred in 24 PCR-positive animals (25.5%), is most closely related to zoonotic B. elizabethae. The remaining three lineages (I-III) formed a monophyletic clade which, on the basis of gltA was shown to contain bartonellae from diverse Gerbillinae species from the Middle East, suggestive of a gerbil-associated species complex in this region. Lineage I was identical to a Candidatus B. sanaae strain identified previously in Bushy-tailed jirds (Sekeetamys calurus) from Egypt, wherease MLSA indicate that lineages II and III are novel. The high levels of infection and co-infection, together with the presence of multiple Bartonella lineages indicate that Gerbillus nanus is likely a natural reservoir of Bartonella in the Arabian Peninsula.

Seroprevalence of Bartonella spp., Coxiella burnetii, and Hantavirus among people who inject drugs in Rio de Janeiro, Brazil: a retrospective assessment of a biobank.

The increasing use of illicit drugs imposes a public health challenge worldwide. People who inject drugs (PWID) are more susceptible to health complications due to immunosuppression associated with drug use and non-hygienic self-administration of substances, contaminants, and liquids. PWID are subjected to increased risk of acquiring and transmitting different pathogens (frequently functioning as sentinel cases for (re)emerging pathogens), including those transmitted by arthropods and vertebrate reservoirs in unhealthy environments. A clear association between injection drug use and HIV, HBV, and HCV infections has been described; however, other infectious viral and bacterial agents have been seldomly assessed. In this study, we investigated the seroprevalence of Bartonella spp., Coxiella burnetii, and Hantavirus among 300 randomly selected PWIDs from Rio de Janeiro, as part of a multi-city cross-sectional study carried out in the 1990s. Point seroprevalences and respective 95% CIs are as follows: 9.3% for C. burnetii (95% CI: 6.0%-13.0%), 1.0% for Bartonella spp. (95% CI: 0.0%-3.0%), and 4.0% for Hantavirus (95% CI: 2.0%-7.0%). In addition to the blood-borne pathogens, the results of this study increase our knowledge on other transmissible infectious agents in PWID. The high seroprevalence of C. burnetii and Hantavirus found among PWID is intriguing and suggests the need to carry out prospective studies, including molecular analyses, to confirm these findings and allow a better understanding of the putative relevance of these zoonotic infectious agents among PWID.

Assessment of a quantitative 5' nuclease real-time polymerase chain reaction using the nicotinamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) for Bartonella species in domiciled and stray cats in Brazil.

OBJECTIVES: The objective of this study was to develop a quantitative 5' nuclease real-time polymerase chain reaction (PCR) assay to diagnose infections caused by Bartonella species. METHODS: Between January and April 2013 whole blood samples were collected by convenience from 151 cats (86 domiciled and 65 stray cats). The feline blood samples were subjected to a novel quantitative 5' nuclease real-time PCR (qPCR) for Bartonella species targeting the nictonamide adenine dinucleotide dehydrogenase gamma subunit (nuoG) and conventional PCR assays targeting intergenic transcribed spacer, ribC, gltA, pap31 and rpoB, followed by sequencing and basic local alignment search tool analysis. RESULTS: The qPCR assay detected as few as 10 copies of plasmid per reaction. Forty-six (54.4% domiciled and 45.6% stray cats) of 151 sampled cats showed positive results in nuoG qPCR for Bartonella species. The absolute quantification of nuoG Bartonella DNA in sampled cats ranged from 1.1 × 10(4) to 1.3 × 10(4). Eighteen (39.1%) of 46 positive samples in the qPCR were also positive in conventional PCR assays. The sequencing confirmed that Bartonella henselae and Bartonella clarridgeiae circulate in cats in midwestern Brazil. CONCLUSIONS AND RELEVANCE: The present work provides details of a novel qPCR assay to diagnose infections caused by Bartonella species.

A multi-gene analysis of diversity of Bartonella detected in fleas from Algeria.

We report the molecular detection of several Bartonella species in 44 (21.5%) of 204 fleas from Algeria collected from 26 rodents and 7 hedgehogs. Bartonella elizabethae and B. clarridgeiae were detected in the fleas collected on hedgehogs. Bartonella tribocorum and B. elizabethae were detected in fleas collected from rats and mice, and sequences similar to an unnamed Bartonella sp. detected in rodents from China were detected in rats as well as a genotype of Bartonella closely related to Bartonella rochalimae detected in fleas collected on brown rats (Rattus norvegicus).

Analysis of multi-strain Bartonella pathogens in natural host population--do they behave as species or minor genetic variants?

Modern advances in genetic analysis have made it feasible to ascertain the variant type of a pathogen infecting a host. Classification of pathogen variants is commonly performed by clustering analysis of the observed genetic divergence among the variants. A natural question arises whether the genetically distinct variants are epidemiologically distinct. A broader question is whether the different variants constitute separate microbial species or represent minor variations of the same species. These important issues were addressed in the context of analyzing dynamics of genetically distinct variants of Bartonella bacteria in cotton rat hosts. Frequencies of acquiring a new variant were measured in relation to the genetic differences between variants successively infecting an individual rodent host. Two statistical techniques were introduced for performing such analysis, and the methodologies were illustrated with a set of data collected from a particular multi-strain Bartonella system. We carried out a frequency analysis of co-infection patterns, and a Markov chain analysis of panels of successive mixed infection time series for testing some particular gene-based grouping of the Bartonella variants with a panel of observed disease data from a rodent population. Our analysis suggests that the three genogroups A, B and C of Bartonella function as independent species but the variants within each genogroup enjoy some cross-immunity against each other. The newly developed methodologies are broadly applicable for analyzing other multi-strain pathogen data which are increasingly collected for diverse infectious diseases.

Rapid and cost-effective identification of Bartonella species using mass spectrometry.

Bacteria of the genus Bartonella are emerging zoonotic bacteria recognized in a variety of human diseases. Due to their poor chemical reactivity, these fastidious bacteria are poorly characterized using routine phenotypic laboratory tests. Identification is usually achieved using molecular techniques that are time-consuming, expensive and technically demanding. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a new technique for bacterial species identification. This study evaluated the use of MALDI-TOF MS for rapid genus and species identification of Bartonella species. Reference strains representing 17 recognized Bartonella species were studied. For each species, MS spectra for four colonies were analysed. The consensus spectrum obtained for each species was unique among spectra obtained for 2843 bacteria within the Bruker database, including 109 alphaproteobacteria. Thirty-nine additional blind-coded Bartonella strains were correctly identified at the species level, including 36 with a significant score. Altogether, these data demonstrate that MS is an accurate and reproducible tool for rapid and inexpensive identification of Bartonella species.

Codon and amino acid usage in two major human pathogens of genus Bartonella--optimization between replicational-transcriptional selection, translational control and cost minimization.

Intra-genomic variation in synonymous codon and amino acid usage in two human pathogens Bartonella henselae and B. quintana has been carried out through multivariate analysis. Asymmetric mutational bias, coupled with replicational-transcriptional selection, has been identified as the prime selection force behind synonymous codon selection--a characteristic of the genus Bartonella, not exhibited by any other alpha-proteobacterial genome. Distinct codon usage patterns and low synonymous divergence values between orthologous sequences of highly expressed genes from the two Bartonella species indicate that there exists a residual intra-strand synonymous codon bias in the highly expressed genes, possibly operating at the level of translation. In the case of amino acid usage, the mean hydropathy level and aromaticity are the major sources of variation, both having nearly equal impact, while strand-specific mutational pressure and gene expressivity strongly influence the inter-strand variations. In both species under study, the highly expressed gene products tend not to contain heavy and/or aromatic residues, following the cost-minimization hypothesis in spite of their intracellular lifestyle. The codon and amino acid usage in these two human pathogens are, therefore, consequences of a complex balance between replicational-transcriptional selection, translational control, protein hydropathy and cost minimization.