Abstract Rift Valley fever virus (RVFV) is a zoonotic bunyavirus that causes abortions in domesticated ruminants. Sheep breeds exotic to endemic areas are reportedly the most susceptible to RVFV infection. Within the scope of a risk assessment program of The Netherlands, we investigated the susceptibility of a native breed of gestating sheep to RVFV infection. Ewes were infected experimentally during the first, second, or third trimester of gestation. Mortality was high among ewes that developed viremia. Four of 11 inoculated ewes, however, did not develop detectable viremia nor clinical signs and did not seroconvert for immunoglobulin G (IgG) or IgM antibodies. Surprisingly, these ewes were found to contain viral RNA in maternal and fetal organs, and the presence of live virus in fetal organs was demonstrated by virus isolation. We demonstrate that RVFV can be transmitted vertically in the absence of detectable maternal viremia.

Abstract The flea-borne rickettsioses murine typhus (Rickettsia typhi) and flea-borne spotted fever (FBSF) (Rickettsia felis) are febrile diseases distributed among humans worldwide. Murine typhus has been known to be endemic to Kenya since the 1950s, but FBSF was only recently documented in northeastern (2010) and western (2012) Kenya. To characterize the potential exposure of humans in Kenya to flea-borne rickettsioses, a total of 330 fleas (134 pools) including 5 species (Xenopsylla cheopis, Ctenocephalides felis, Ctenocephalides canis, Pulex irritans, and Echidnophaga gallinacea) were collected from domestic and peridomestic animals and from human dwellings within Asembo, western Kenya. DNA was extracted from the 134 pooled flea samples and 89 (66.4%) pools tested positively for rickettsial DNA by 2 genus-specific quantitative real-time PCR (qPCR) assays based upon the citrate synthase (gltA) and 17-kD antigen genes and the Rfelis qPCR assay. Sequences from the 17-kD antigen gene, the outer membrane protein (omp)B, and 2 R. felis plasmid genes (pRF and pRFd) of 12 selected rickettsia-positive samples revealed a unique Rickettsia sp. (n=11) and R. felis (n=1). Depiction of the new rickettsia by multilocus sequence typing (MLST) targeting the 16S rRNA (rrs), 17-kD antigen gene, gltA, ompA, ompB, and surface cell antigen 4 (sca4), shows that it is most closely related to R. felis but genetically dissimilar enough to be considered a separate species provisionally named Candidatus Rickettsia asemboensis. Subsequently, 81 of the 134 (60.4%) flea pools tested positively for Candidatus Rickettsia asemboensis by a newly developed agent-specific qPCR assay, Rasemb. R. felis was identified in 9 of the 134 (6.7%) flea pools, and R. typhi the causative agent of murine typhus was not detected in any of 78 rickettsia-positive pools assessed using a species-specific qPCR assay, Rtyph. Two pools were found to contain both R. felis and Candidatus Rickettsia asemboensis DNA and 1 pool contained an agent, which is potentially new.

Abstract

Background:

Leptospirosis and human immunodeficiency virus (HIV) infection are prevalent in many areas, including northern Tanzania, yet little is known about their interaction.

Methods:

We enrolled febrile inpatients at two hospitals in Moshi, Tanzania, over 1 year and performed HIV antibody testing and the microscopic agglutination test (MAT) for leptospirosis. Confirmed leptospirosis was defined as ≥four-fold rise in MAT titer between acute and convalescent serum samples, and probable leptospirosis was defined as any reciprocal MAT titer ≥800.

Results:

Confirmed or probable leptospirosis was found in 70 (8.4%) of 831 participants with at least one serum sample tested. At total of 823 (99.0%) of 831 participants had HIV testing performed, and 203 (24.7%) were HIV infected. Among HIV-infected participants, 9 (4.4%) of 203 had confirmed or probable leptospirosis, whereas among HIV-uninfected participants 61 (9.8%) of 620 had leptospirosis. Leptospirosis was less prevalent among HIV-infected as compared to HIV-uninfected participants [odds ratio (OR) 0.43, p=0.019]. Among those with leptospirosis, HIV-infected patients more commonly presented with features of severe sepsis syndrome than HIV-uninfected patients, but differences were not statistically significant. Among HIV-infected patients, severe immunosuppression was not significantly different between those with and without leptospirosis (p=0.476). Among HIV-infected adolescents and adults, median CD4 percent and median CD4 count were higher among those with leptospirosis as compared to those with other etiologies of febrile illness, but differences in CD4 count did not reach statistical significance (p=0.015 and p=0.089, respectively).

Conclusions:

Among febrile inpatients in northern Tanzania, leptospirosis was not more prevalent among HIV-infected patients. Although some indicators of leptospirosis severity were more common among HIV-infected patients, a statistically significant difference was not demonstrated. Among HIV-infected patients, those with leptospirosis were not more immunosuppressed relative to those with other etiologies of febrile illness.

Abstract Although Crimean-Congo hemorrhagic fever (CCHF) is a widespread tick-borne disease, little is known about its pathogenesis. The interaction of the virus with host cells is most likely responsible for the pathogenesis of CCHF. The main contributors are endothelial cells (ECs) and immune cells. There are 2 theories underlying the CCHF pathogenesis: One is that the virus interacts with the ECs directly and the other that it interacts indirectly via immune cells with subsequent release of soluble mediators. ECs are activated upon infection by the upregulation of soluble molecules and proinflammatory cytokines. Probably, in severe cases, deregulation and excessive release of the cytokines accompanied by endothelial activation have toxic effects, leading to increased vascular permeability, vasodilatation, and subsequently hypotension, multiple organ failure, shock, and death. Studies indicate that CCHF virus (CCHFV) also can impair the innate immune system and cause a delay in adaptive immune response, which is critical for the clearance of CCHFV. The virus has many different ways to block the immune response, leading to uncontrolled viral replication followed by systemic spread of the virus throughout the body. Partial activation of dendritic cells and macrophages, delayed induction of interferons, weak antibody response, apoptosis of lymphocytes, and hemophagocytosis are some of these tactics. However, there are many points waiting for clarification about the pathogenesis of CCHF. Although the high risk of contagiousness limits research, we need more studies to understand the CCHF pathogenesis better. Here we review the main characteristics of the pathogenesis of CCHF.

Abstract Introduction: Rickettsia felis is an emerging agent considered a human threat; although its natural reservoir and agent of transmission is the cat flea, it has been also found in other vectors. R. felis has been identified in Chile in cat fleas and in one specimen of Rhipicephalus sanguineus collected in the Metropolitan Region. The objective of this study was to detect the presence of Rickettsia spp. in R. sanguineus from dogs of two different and distant geographical areas in Chile. Material and

Methods:

We performed a domiciliary sampling in urban and rural localities of two distant areas of the country-the Metropolitan Region in the center and the northern city of Arica. A total of 460 households were visited; one dog per household was included in the study and ectoparasites were collected from them.

Results:

R. sanguineus was found in 50% of the 460 dogs. R. felis was identified by amplification and sequencing of gltA, ompA, and ompB genes in R. sanguineus from both regions, with predominance in Arica. Discussion: The presence of R. felis in R. sanguineus from two distant regions of Chile suggests that this rickettsial agent is well established in the country. Considering that no human spotted fever group infections have been recognized in the country, the results should alert clinicians about such possible cases. The role of R. sanguineus in the epidemiology and transmission of R. felis should be further investigated.

Abstract Chlamydia psittaci is a zoonotic pathogen with a wide range of avian hosts and worldwide geographical spread. Zoonotic transfer occurs by inhalation or direct contact and may cause psittacosis or parrot disease. Host reservoirs of particular epidemiological interest include gregarious or migratory species, as colonial behavior facilitates microbial spread amongst conspecifics and a migratory ecology permits disperse over a wide geographic region. The current study detected C. psittaci antibodies in 76 of 81 (93.8%) feral Canada geese (Branta canadensis) using a species-specific enzyme-linked immunosorbent assay. Viable C. psittaci were isolated from pharyngeal swabs of 47 of 81 (58%) birds, and subsequent ompA-based genotyping revealed genotypes E, B, and E/B. Absence of clinical symptoms in these geese could reflect a strong natural protection or a persistent infection. Canada geese are considered an important alien species in Europe. Current results denote that this avian reservoir poses a considerable risk to native wildlife as a potential source of C. psittaci.

Abstract Highly pathogenic avian influenza virus A/H5N1 has been reported in 11 African countries. Migratory waterbirds have the potential of introducing A/H5N1 into east Africa through the Rift Valley of Kenya. We present the results of a wild bird surveillance system for A/H5N1 and other avian influenza viruses based on avian fecal sampling in Kenya. We collected 2630 fecal samples in 2008. Viral RNA was extracted from pools of 3-5 fecal samples and analyzed for presence of avian influenza virus RNA by real-time RT-PCR. Twelve (2.3%) of the 516 sample pools were positive for avian influenza virus RNA, 2 of which were subtyped as H4N6 viruses. This is the first report of avian influenza virus in wild birds in Kenya. This study demonstrates the success of this approach in detecting avian influenza virus in wild birds and represents an efficient surveillance system for avian influenza virus in regions with limited resources.

Abstract

Background:

Tularemia, caused by a Gram-negative bacterium Francisella tularensis, is an occasional disease of cats in the midwestern United States and a public health concern due to its zoonotic potential. Different environmental, climatic, and pet-owner's housing and socioeconomic conditions were evaluated as potential risk factors for feline tularemia using Geographic Information Systems (GIS) in a retrospective case-control study.

Methods:

The study included 46 cases identified as positive for tularemia based upon positive immunohistochemistry, isolation of F. tularensis using bacterial culture, and 4-fold or greater change in serum antibody titer for F. tularensis. Cats with a history of fever, malaise, icterus, and anorexia but no lesions characteristic of tularemia and/or negative immunohistochemistry, no isolation of bacteria in bacterial culture, and less than 4-fold raise in serum antibody titer for F. tularensis were treated as controls (n=93). Candidate geospatial variables from multiple thematic sources were analyzed for association with case status. Variables from National Land Cover Dataset, Soil Survey Geographic Database, US Census Bureau, and Daymet were extracted surrounding geocoded case-control household locations. Univariable screening of candidate variables followed by stepwise multivariable logistic modeling and odds ratios were used to identify strengths of variable associations and risk factors.

Results:

Living in a residence located in newly urbanized/suburban areas, residences surrounded by areas dominated by grassland vegetation, and mean vapor pressure conditions recorded during the 8(th) week prior to case arrival at the hospital are significant risk factors for feline tularemia.

Conclusions:

Prevention strategies such as acaricide applications in residential backyards during spring and early summer periods and any behavior modifications suitable for cats that will prevent them from contracting infection from ticks or dead animals are necessary. Mean vapor pressure conditions recorded during the 8(th) week prior to case arrival at a diagnostic facility is a predictor for feline tularemia.

Abstract Orientia tsutsugamushi, the etiologic agent of potentially fatal scrub typhus, is characterized by a high antigenic diversity, which complicates the development of a broadly protective vaccine. Efficacy studies in murine and nonhuman primate models demonstrated the DNA vaccine candidate pKarp47, based upon the O. tsutsugamushi Karp 47-kD HtrA protein gene, to be a successful immunoprophylactic against scrub typhus. To characterize 47-kD HtrA protein diversity among human isolates of Orientia, we sequenced the full open reading frame (ORF) of the 47-kD HtrA gene and analyzed the translated amino acid sequences of 17 patient isolates from Thailand (n=13), Laos (n=2), Australia (n=1), and the United Arab Emirates (UAE) (n=1) and 9 reference strains: Karp (New Guinea), Kato (Japan), Ikeda (Japan), Gilliam (Burma), Boryong (Korea), TA763, TH1811 and TH1817 (Thailand), and MAK243 (China). The percentage identity (similarity) of translated amino acid sequences between 16 new isolates and 9 reference strains of O. tsutsugamushi ranged from 96.4% to 100% (97.4% to 100%). However, inclusion of the recently identified Orientia chuto sp. nov. reduced identity (similarity) values to 82.2% to 83.3% (90.4% to 91.4%). These results demonstrate the diversity of Orientia 47-kD HtrA among isolates encountered by humans and therefore provide support for the necessity of developing a broadly protective scrub typhus vaccine that takes this diversity into account.