Development and validation of a triplex real-time qPCR for sensitive detection and quantification of major rat bite fever pathogen Streptobacillus moniliformis.

Streptobacillus (S.) moniliformis is the most important pathogen causing rat bite fever (RBF) worldwide. This zoonotic pathogen is understudied mainly due to difficulties in culturing S. moniliformis as a fastidious microorganism. Therefore, advances in molecular detection techniques are highly needed, especially with regard to the widespread availability of real-time quantitative (q) PCR in laboratories. In this study, we aimed to develop a qPCR for the identification of Streptobacillus species and quantification of S. moniliformis in clinical samples, especially those derived from tissue samples of animal origin. We optimized a previously described PCR protocol in order to develop a qPCR, which can detect different Streptobacillus species with high specificity and is simultaneously able to quantitate S. moniliformis in different clinical matrices. The qPCR exhibited a limit of detection (LOD) of 21 copies/reaction representing ~4-5 streptobacilli, while the limit of quantification (LOQ) was 2.1 × 103 copies/reaction. It was also more sensitive than conventional PCR by two orders of magnitude and proved to have a substantial agreement (Kappa 0.74) compared to it with a superior detection rate in 374 samples from wild rats, laboratory rats and animals from holdings of wild-trapped rats. To conclude, the qPCR described in this study is an important molecular tool that is able to quantify S. moniliformis in tissue samples of animal origin. It represents a suitable tool for future establishment and evaluation of other molecular assays that are highly needed for a better understanding of epidemiology and pathophysiology of RBF. In experimental studies, it will also be useful for titration purposes since the quantification of the organism using classical plate counting technique is problematic and inaccurate.

Rat bite fever: a case report review.

CONTEXT: Rat bite fever is a rare disease with a challenging differential diagnosis. The zoonosis has a potentially lethal course in a vulnerable population (children and low socioeconomic class) and a commonly available standard therapy (penicillin). This case report review outlines common epidemiological and clinical factors to improve clinical awareness and timely response to therapeutic actions. METHODS: A systematic literature review was conducted in the PubMed database looking for English language European case reports of rat bite fever from 2000 to 2021. RESULTS: In 17 out of 20 selected cases, the condition of the index patient was identified as an infectious syndrome. Thanks to the almost omnisensitive susceptibility pattern of Streptobacillus moniliformis, timely antibiotic administration prevented an unfavorable outcome in all these cases. However, in the three remaining cases, the initial diagnoses were arthritis (on autoimmune basis and gout) and viral syndrome. Due to delayed antibiotic administration, one case suffered persistent harm, while the other two cases encountered prolonged illness. CONCLUSION AND RECOMMENDATIONS: Rat bite fever is a diagnosis that can be easily missed from both a clinical and a microbiological point of view. As such, rat bite fever becomes part of the differential diagnosis whenever a patient presents with a fever syndrome after being in contact with rodents. In the case of persistent fever, blood culture sampling should be performed even in the absence of a systemic inflammatory response. A bacterial 16S ribosomal RNA PCR on blood or joint aspiration (cultures) is an even more sensitive diagnostic test. Since most transmissions occurred in a domestic setting, keeping rats as pets cannot be recommended.

Rat bite fever due to Streptobacillus notomytis complicated by meningitis and spondylodiscitis: a case report.

BACKGROUND: Only three other cases of rat bite fever caused by Streptobacillus notomytis in humans have been reported since this species was identified in 2015. Data specific to the differences in clinical features and geographic distribution between S. notomytis infection and S. moniliformis infection are scarce. All previous cases of human S. notomytis infection were reported from Japan. This is the first case of S. notomytis infection reported from outside of Japan. CASE PRESENTATION: A 72-year-old Thai woman was admitted to Siriraj Hospital (Bangkok, Thailand)-Thailand's largest university-based national tertiary referral center-in August 2020 with fever, myalgia, and polyarthralgia for 3 days, and gradually decreased consciousness for the past 1 day. Physical examination and laboratory investigations revealed septic arthritis of both knee joints, meningitis, and hepatitis. She was initially misdiagnosed as rheumatoid arthritis in the elderly since the initial investigations were unable to detect a causative pathogen. However, S. notomytis infection was later confirmed by polymerase chain reaction amplification of a part of the 16S rRNA gene and sequencing from synovial fluid. Her clinical course was also complicated by spondylodiscitis and epidural abscess caused by S. notomytis, which was detected from tissue biopsy. Therefore, rat bite fever in this patient manifested as meningitis, septic polyarthritis, hepatitis, and spondylodiscitis. The patient was treated with intravenous ceftriaxone then switched to oral amoxicillin with complete recovery. CONCLUSIONS: The clinical manifestations of S. notomytis infection are similar to those demonstrated in S. moniliformis infection. This case also showed that arthritis caused by S. notomytis mimics rheumatoid arthritis, and that meningitis and spondylodiscitis are potential coexisting complications that can be found in S. notomytis infection.

Prevalence and Diversity of the Streptobacillus Rat-bite Fever Agent, in Three Invasive, Commensal Rattus Species from South Africa.

Rat-bite fever is an over-looked, global zoonotic disease that has a mortality rate of up to 13%, if untreated. Historically, this rat-borne disease has been attributed to one of two causative agents, Streptobacillus moniliformis or Spirillum minus. Given the confirmed presence of multiple invasive Rattus host species, high rat densities in urban, informal human settlements and increasing reports of rat bites in South Africa, we undertook a retrospective assessment of Streptobacillus in rats sampled from 16 urban sites, in Gauteng, the smallest but most populous Province in South Africa. Using a multi-gene PCR-sequencing approach, we confirmed Streptobacillus presence in 50.9% of oral swabs from three rat species and the presence of two Streptobacillus species, viz.S. moniliformis and S. notomytis. The two members of the cryptic Rattus rattus species complex (R. rattus and R. tanezumi), which are morphologically indistinguishable from each other, had markedly different colonization rates. Whereas 48.6% of rats from this species complex were Streptobacillus-positive, only 32.3% of Rattus tanezumi were positive compared to 61.5% R. rattus. Rattus norvegicus had an intermediate prevalence of 55.6%. Phylogenetic analysis of four gene regions (16S rRNA, gyrB, groEL, recA) identified two discrete lineages; S. moniliformis occurred exclusively in R.norvegicus, and S. notomytis was restricted to the two members of the R. rattus species complex; this represents the first report of Streptobacillus in R. tanezumi. These results highlight a largely overlooked zoonotic threat posed by invasive rats and confirm the presence of two discrete and potentially host-specific Streptobacillus lineages in South Africa.

Erosive polyarthritis caused by sepsis due to a novel species of Streptobacillus notomytis.

Rat-bite fever (RBF) is characterized by fever, rash and arthritis, mainly caused by Streptobacillus moniliformis. We present a case of inflammatory erosive polyarthritis with sepsis caused by Streptobacillus notomytis, a novel species isolated from S. moniliformis. A 67-year-old man presented with fever, purpura and peripheral arthritis. After blood cultures were performed, loxoprofen administration was initiated. Arthritis partially improved, but left first metatarsophalangeal (MTP) arthritis persisted. A gram-negative rod was detected in the blood culture, and meropenem administration was started that improved arthritis. Ultrasonography and computed tomography revealed bone erosion in the left first MTP joint, leading to the diagnosis of septic arthritis. 16S rRNA gene sequence analysis revealed that this strain was S. notomytis. Further questioning revealed the patient may have had contact with rats. It is essential to consider Streptobacillus infection in the differential diagnosis of erosive polyarthritis.

Streptobacillus felis, a member of the oropharynx microbiota of the Felidae, isolated from a tropical rusty-spotted cat.

Streptobacillus felis is a fastidious microorganism and a novel member of the potentially zoonotic bacteria causing rat bite fever. Since its description, this is the second isolation of S. felis in a diseased member of the Felidae. Interestingly, the strain from this study was isolated from a zoo held, rusty-spotted cat (Prionailurus rubiginosus), with pneumonia, thereby indicating a possible broader host range in feline species. A recent preliminary sampling of domestic cats (Felis silvestris forma catus) revealed that this microorganism is common in the oropharynx, suggesting that S. felis is a member of their normal microbiota. Due to unawareness, fastidiousness, antibiotic sensitivity and lack of diagnostics the role of S. felis as a cat and human pathogen might be under-reported as with other Streptobacillus infections. More studies are necessary to elucidate the role of S. felis in domestic cats and other Felidae in order to better estimate its zoonotic potential.

Rat bite fever in senior health medicine.

A 71-year-old man was admitted with confusion having been discovered on the floor of his unkempt home. Social services would later confirm that the home was infested by rats. An infection of unclear origin was suspected, and he was immediately started on broad spectrum antibiotics. He subsequently developed a swollen left knee. Two knee aspirates showed a raised white cell count but no microbiological or cytological diagnosis. Finally, samples sent for 16S rDNA PCR identified Streptobacillus monilliformis Fastidious organisms and early treatment with antibiotics can limit microbiological diagnosis through standard laboratory analysis. Newer techniques such as sequence-based testing can overcome these limitations; however, they are not widely available and require the clinician to have a high index of suspicion. Crucially, the patient continued to be treated for his clinical sepsis despite initial laboratory results being negative and conflicting specialist opinions.

Rat bite fever caused by Streptobacillus moniliformis infection in a Chinese patient.

BACKGROUND: Rat bite fever (RBF), a severe infectious disease, can result from transmission of the pathogen Streptobacillus moniliformis (S. moniliformis) by rat bite. RBF diagnosis can be overlooked. CASE PRESENTATION: We present a case of RBF in a Chinese patient who was infected with S. moniliformis in mainland China. Meta-next generation sequencing (mNGS) was used to identify potential pathogens and detected S. moniliformis genome sequences in the pustular sample in less than 72 h. Then the diagnosis was validated by polymerase chain reaction analysis. Despite having severe RBF with complications, this 54-year-old male patient was successfully cured with penicillin as a result of timely pathogen-based diagnosis. CONCLUSIONS: Physicians should inquire about recent rat exposure and consider the possibility of RBF when a patient develops unexplained fever and rashes. mNGS is a new diagnostic technology and may identify RBF pathogens even when blood culture results are negative.

Rat-Bite Fever in Human with Streptobacillus notomytis Infection, Japan.

We report a case of rat-bite fever in a 94-year-old woman with Streptobacillus notomytis infection. We established an epidemiologic link between exposure to rats and human infection by performing nested PCRs that detected S. notomytis in the intraoral swab specimens obtained from rats captured in the patient's house.

Clinical infection in house rats (Rattus rattus) caused by Streptobacillus notomytis.

Rat bite fever is an under-reported, under-diagnosed emerging zoonosis with worldwide distribution. Besides Spirillum minus, Streptobacillus moniliformis is the major causative microorganism although it usually colonises rats without any clinical signs. A group of house rats (Rattus rattus) kept in a zoo exhibition for educational purposes suffered from neurological signs including disorientation, torticollis, stall walking, ataxia and death. Gross pathological and histo-pathological examinations of the investigated rats revealed high-grade otitis interna et media, from which Streptobacillus notomytis was isolated in pure culture or as the predominant microorganism. This case series underlines a previously expressed hypothesis that R. rattus might be naturally colonised with S. notomytis, whereas the traditional rat bite fever organism, S. moniliformis, might be restricted to the Norway rat (Rattus norvegicus). However, the general paucity of Streptobacillus isolates, especially from their respective animal hosts, precludes definitive proof of these host tropisms. This is the first report of S. notomytis detection outside Asia and Australia and the first evidence for its role as a facultative pathogen in house rats.