Prospective serological and molecular cross-sectional study focusing on Bartonella and other blood-borne organisms in cats from Catalonia (Spain)

Background There is limited clinical or epidemiological knowledge regarding Bartonella infection in cats, and no serological studies have compared the presence of antibodies against different Bartonella species. Moreover, there are limited feline Bartonella studies investigating co-infections with other vector-borne pathogens and the associated risk factors. Therefore, the objective of this study was to investigate Bartonella spp. infections and co-infections with other pathogens in cats from Barcelona (Spain) based on serological and/or molecular techniques and to determine associated risk factors. Methods We studied colony and owned cats (n = 135). Sera were tested for Bartonella henselae-, Bartonella quintana-, and Bartonella koehlerae-specific antibodies using endpoint in-house immunofluorescence antibody assays. Bartonella real-time PCR (qPCR) and conventional PCR (cPCR) were performed. In addition, cPCR followed by DNA sequencing was performed for other pathogenic organisms (Anaplasma, Babesia, Cytauxzoon, Ehrlichia, Hepatozoon, hemotropic Mycoplasma, and Theileria spp.). Results From 135 cats studied, 80.7% were seroreactive against at least one Bartonella species. Bartonella quintana, B. koehlerae, and B. henselae seroreactivity was 67.4, 77.0, and 80.7%, respectively. Substantial to almost perfect serological agreement was found between the three Bartonella species. Colony cats were more likely to be Bartonella spp.-seroreactive than owned cats. Moreover, cats aged ≤ 2 years were more likely to be Bartonella spp.-seroreactive. Bartonella spp. DNA was detected in the blood of 11.9% (n = 16) of cats. Cats were infected with B. henselae (n = 12), B. clarridgeiae (n = 3), and B. koehlerae (n = 1). Mycoplasma spp. DNA was amplified from 14% (n = 19) of cat blood specimens. Cats were infected with Mycoplasma haemofelis (n = 8), Candidatus M. haemominutum (n = 6), Candidatus Mycoplasma turicensis (n = 4), and Mycoplasma wenyonii (n = 1). Anaplasma, Babesia, Cytauxzoon, Ehrlichia spp., Hepatozoon, and Theileria spp. DNA was not amplified from any blood sample. Of the 16 Bartonella spp.-infected cats based on PCR results, six (37%) were co-infected with Mycoplasma spp. Conclusions Bartonella spp. and hemoplasma infections are prevalent in cats from the Barcelona area, whereas infection with Anaplasma spp., Babesia, Cytauxzoon, Ehrlichia spp., Hepatozoon, and Theileria infections were not detected. Co-infection with hemotropic Mycoplasma appears to be common in Bartonella-infected cats. To our knowledge, this study is the first to document M. wenyonii is infection in cats. Graphical Abstract

Bartonellosi e D-dimero

The paper reports the case of a 17-year-old immunocompetent boy with persistent fever and elevated D-dimer. The family referred contact with kitten and ingestion of homemade deer salami. Abdominal ultrasound scan and CT showed multiple hepatic and splenic abscesses. High levels of antibody title for Bartonella henselae confirmed the diagnosis of hepatosplenic bartonellosis.

Eritema nodoso: gatta ci cova

Erythema nodosum is a panniculitis that can be triggered by many different stimuli. The paper describes the case of a child who presented with erythema nodosum as the unique clinical manifestation of cat scratch disease. Bartonella henselae infection usually presents with non-tender papule in the scratch line followed by subsequent onset of regional lymphadenopathy eventually associated with systemic symptoms. It can also present with atypical manifestation, such as erythema nodosum. The heterogeneity of the clinical presentations makes the disease to be underdiagnosed, whereby it is important to recognize atypical manifestations. Therefore, it is recommended to include Bartonella henselae serology in the diagnostic evaluation of erythema nodosum.

Diagnosis Based on History, Not Examination

Cat scratch disease (CSD), caused by the bacterium Bartonella henselae, is usually self-limiting, presenting with low-grade fever and tender lymphadenopathy. With delayed diagnosis or reactivation of latent disease, CSD is associated with severe debilitating symptoms. We are presenting a patient who proved adiagnostic challenge, before being found to have reactivation of Bartonella henselae, requiring three-months IV Trimethoprim-Sulfamethoxazole andHydrocortisone.

The first report of the seroprevalence of antibodies against Bartonella spp. in water buffaloes (Bubalus bubalis) from South Thailand

Background and Aim: Bartonellosis is an emerging worldwide zoonosis caused by bacteria belonging to the genus Bartonella. Several studies have been conducted on the prevalence of Bartonella infections from animals and humans, including reports from wild and domestic ruminants. However, there has been only one report of Bartonella infection in water buffaloes from the northeastern part of Thailand. Moreover, the seroprevalence of Bartonella spp. in water buffaloes still remains unknown. This study was conducted to explore the prevalence of Bartonella spp. among water buffaloes from South Thailand using molecular and serological techniques. Materials and Methods: A total of 312 samples (156 blood and 156 sera) of 156 water buffaloes from 29 farms in Phatthalung Province, South Thailand, were collected from January to March 2021. All samples were screened for Bartonella spp. using polymerase chain reaction and indirect immunofluorescence assay. Results: The seroprevalence of antibodies against three Bartonella spp. was 16.03% (25/156, 95% confidence interval: 10.65-22.74%), and among 25 water buffaloes with seroprevalence, 56%, 20%, and 24% were positive for antibodies against Bartonella henselae, Bartonella vinsonii subspp. berkhoffii, and Bartonella tamiae, respectively. No significant difference was detected among seroprevalence, gender, age, and ectoparasite infestation. Conclusion: This is the first report of the seroprevalence of antibodies against B. henselae, B. vinsonii subspp. berkhoffii, and B. tamiae in water buffaloes from South Thailand. Further studies are required on the epidemiology of Bartonella infection among water buffaloes, related personnel, and ectoparasites.

Investigation of Transovarial Transmission of Bartonella henselae in Rhipicephalus sanguineus sensu lato Ticks Using Artificial Feeding

Bartonella henselae is a slow-growing, Gram-negative bacterium that causes cat scratch disease in humans. A transstadial transmission of the bacteria from larvae to nymphs of Rhipicephalus sanguineus sensu lato (s.l.) ticks, suspected to be a potential vector of the bacteria, has been previously demonstrated. The present study aims to investigate transovarial transmission of B. henselae from R. sanguineus s.l. adults to their instars. Adult ticks (25 males and 25 females) were fed through an artificial feeding system on B. henselae-infected goat blood for 14 days, and 300 larvae derived from the experimentally B. henselae-infected females were fed on noninfected goat blood for 7 days. Nested PCR and culture were used to detect and isolate B. henselae in ticks and blood samples. Bartonella henselae DNA was detected in midguts, salivary glands, and carcasses of the semi-engorged adults and pooled tick feces (during feeding and post-feeding periods). After the oviposition period, B. henselae DNA was detected in salivary glands of females (33.3%), but not in pooled eggs or larvae derived from the infected females. However, B. henselae DNA was detected by nested PCR from the blood sample during larval feeding, while no viable B. henselae was isolated by culture. According to our findings, following infected blood meal, B. henselae could remain in the tick midguts, move to other tissues including salivary glands, and then be shed through tick feces with limited persistency. The presence of bacterial DNA in the blood during larval feeding shows the possibility of transovarial transmission of B. henselae in R. sanguineus s.l. ticks.

СЕРОЛОГІЧНА ДІАГНОСТИКА КЛІЩОВИХ ІНФЕКЦІЙ У ХВОРИХ НА ЛОКАЛІЗОВАНУ СКЛЕРОДЕРМІЮ

Мета дослідження – встановити частоту виявлення специфічних антитіл IgM і/чи IgG до Borrelia burgdorferi s. l., B. miyamotoi, Bartonella henselae та B. quintana у сироватці крові хворих на локалізовану склеродермію. Пацієнти і методи. Під спостереженням було 78 хворих із локалізованою склеродермією, віком від 18 до 74 років, які протягом 2015-2021 рр. лікувались амбулаторно і стаціонарно в КУТОР «Тернопільський обласний клінічний шкірно-венерологічний диспансер». Чоловіків було 17 (21,8 %), жінок – 61 (78,2 %). Для виявлення специфічних IgM і/чи IgG до B. burgdorferi s. l. (збудників Лайм-бореліозу) у сироватці крові використали двохетапний метод (ІФА та імуноблот) за допомогою тест-систем компанії Euroimmun AG (Німеччина). Отримані результати аналізували відповідно до рекомендацій виробника. Антитіла IgM і IgG до B. miyamotoi (одного зі збудників кліщових поворотних гарячок) визначали в сироватці крові методом імуноблоту в лабораторії «IGeneX Inc.» (Мілпітас, Каліфорнія, США). Специфічні антитіла IgG до Bartonella henselae та Bartonella quintana (збудників бартонельозу) визначали у сироватці крові пацієнтів за допомогою методу мультиплексної непрямої імунофлуоресценції, застосувавши тест-системи «Mosaic for Bartonella henselae / Bartonella quintana (IgG)», компанії Euroimmun AG (Німеччина), із використанням технології «Біочіп», які містили мічені флуоресцеїном антигени вказаних видів бартонел. Результати досліджень та їх обговорення. Позитивні або проміжні результати пошуку специфічних IgM і/чи IgG до комплексу B. burgdorferi s. l. (хоча б одного класу антитіл) за допомогою ІФА отримано в 29 (37,2 %) із 78 пацієнтів з локалізованою склеродермією. Підтвердити отримані дані методом імуноблоту вдалося у 25 (86,2 %) хворих. Антитіла лише класу IgM одночасно до B. burgdorferi s. l. та B. miyamotoi методом імуноблоту діагностовано у 4 (11,1 %), IgG у – у 5 (13,9 %) із 36 пацієнтів із локалізованою склеродермією. За допомогою методу непрямої імунофлуоресценції специфічні антитіла класу G лише до B. henselae виявлено в сироватці крові 4 (15,3 %) із 26 пацієнтів із локалізованою склеродермією. Серологічну діагностику кліщових поворотних гарячок і бартонельозу (наявних або перенесених у минулому) в пацієнтів із локалізованою склеродермією, мешканців Тернопільської області, проведено вперше. Висновки. Застосування двохетапного методу серологічної діагностики ЛБ (ELISA та імуноблот) дозволило виявити антитіла IgM і/чи IgG до B. burgdorferi s. l. у 32,1 % хворих із локалізованою склеродермією. Встановлено причетність B. miyamotoi до клінічних проявів локалізованої склеродермії у 13,9 % пацієнтів шляхом виявлення у них антитіл IgG одночасно до B. miyamotoi та B. burgdorferi s. l. методом імуноблоту. Специфічні антитіла IgG лише до B. henselae діагностовано в сироватці крові 15,3 % пацієнтів із локалізованою склеродермією.