Molekularni dokaz filarija u pasa u Nigeriji, zapadna Afrika

The filarioid worm Dirofilaria repens has long been reported in dogs in Nigeria. Recent studies however, did not only report increased prevalence of D. repens but also the presence of Dirofilaria immitis. The classical diagnostic methods used in these studies have low sensitivity. Therefore, we screened 197 canine blood samples from seven states in Nigeria, using a highly sensitive and specific High Resolution Melt Real Time PCR and sequencing, to determine the prevalence and species of filarial worms infecting Nigerian dogs. Only one (0.5%) of the 197 samples screened was positive and showed a melt curve similar to Acanthocheilonema reconditum. Nevertheless, the sequence of this positive sample had only 94% similarity to its first GenBank match, a A. reconditum (JF461460.1). This could be a new filarioid species or a variant of an existing species and deserves further investigation. The low prevalence reported herein is in discrepancy with previous reports that showed the frequent presence of canine filariasis in Nigeria. A large scale survey is needed of filarioids infecting dogs in Nigeria, using highly sensitive and specific methods, to identify the present species and provide a baseline data on their national prevalence and geographic distribution.O parazitu Dirofilaria repens u pasa u Nigeriji odavno postoje podaci. Novija istraživanja, međutim, ne pokazuju samo povećanu prevalenciju D. repens nego i prisutnost D. immitis. Klasične dijagnostičke metode primijenjene u ovom istraživanju imaju nisku osjetljivost. Stoga smo ispitali 197 uzoraka pseće krvi iz sedam pokrajina u Nigeriji koristeći se visokoosjetljivom i visokospecifičnom metodom High Resolution Melt Real Time PCR i sekvenciranjem kako bismo odredili prevalenciju i vrste filarija koje invadiraju pse u Nigeriji. Samo je jedan (0,5 %) od 197 uzoraka bio pozitivan i pokazao je krivulju sličnu krivulji za parazita Acanthocheilonema reconditum. Sekvencija tog pozitivnog uzorka imala je samo 94 % sličnosti s prvom podudarnom sekvencijom iz banke gena, A. reconditum (JF461460.1). To bi mogla biti nova vrsta filarija ili varijanta postojećih vrsta i zahtijeva daljnja istraživanja. Opisana niska prevalencija u ovome radu odstupa od prijašnjih izvješća koja pokazuju čest nalaz psećih filarija u Nigeriji. Kako bi se dokazale postojeće vrste i pružili referentni podaci o prevalenciji i zemljopisnoj raširenosti filarija u pasa u Nigeriji, potrebno je provesti opsežnija istraživanja

Molecular Detection and Identification of Spotted Fever Group Rickettsiae in Ticks Collected from the West Bank, Palestinian Territories

Tick-borne rickettsioses are caused by obligate intracellular bacteria belonging to the spotted fever group (SFG) rickettsiae. Although Spotted Fever is prevalent in the Middle East, no reports for the presence of tick-borne pathogens are available or any studies on the epidemiology of this disease in the West Bank. We aimed to identify the circulating hard tick vectors and genetically characterize SFG Rickettsia species in ixodid ticks from the West Bank-Palestinian territories.This study was funded by the Netherlands Ministry of Foreign Affairs, The Hague, The Netherlands and NVHU under grant reference number 2014.52146 and USAID grant MERC TAMOU- 12-M32-038. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript

Molecular Detection of Bartonella Species in Fleas Collected from Dogs and Cats from Costa Rica

The bacterial genus Bartonella includes several species with zoonotic potential, some of which are common in domestic dogs and cats, as well as in their fleas. Because there is no previous information about the presence of Bartonella species in fleas from Central America, this study aimed at evaluating the presence of Bartonella spp. in fleas collected from dogs and cats in Costa Rica. A total 72 pools of Ctenocephalides felis and 21 pools of Pulex simulans were screened by conventional PCR to detect Bartonella DNA fragments of the citrate synthase (gltA) and the b subunit RNA polymerase (rpoB) genes. Three (4.2%) pools of C. felis and five pools (22.7%) of P. simulans were found positive for Bartonella DNA. Sequences corresponding to Bartonella vinsonii subsp. berkhoffii strain Winnie, B. rochalimae, and an undescribed Bartonella sp. (clone BR10) were detected in flea pools from dogs, whereas Bartonella henselae and B. clarridgeiae sequences were identified in flea pools from cats. The detection of zoonotic Bartonella spp. in this study should increase the awareness to these flea borne diseases among physicians and public health workers and highlight the importance of flea control in the region.Universidad de Costa Rica/[803-A8-127]/UCR/Costa RicaRed para la investigación y el entrenamiento en enfermedades tropicales/[9N-2008]/NETROPICA/Costa RicaUCR::Vicerrectoría de Docencia::Salud::Facultad de MicrobiologíaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET

Guidelines for the isolation, molecular detection, and characterization of Bartonella Species

Bartonellae are fastidious, facultative, intracellular vector-borne bacteria distributed among mammalian reservoirs worldwide. The pathogenic potential of many Bartonella spp. has increased the interest in these bacteria and advanced their research. Isolation of Bartonella spp. is laborious using classical bacteriological methods and requires specific conditions and prolonged incubation periods. In contrast, molecular methods for detection of Bartonella DNA are considered as more practical and sensitive than the former. Among the molecular methods, the use of real-time PCR assays for primary screening of Bartonella spp., followed by several molecular confirmatory assays, using either conventional or real-time PCR, is recommended. Although primary isolation of Bartonella is a laborious task, we encourage its application to all PCR-positive samples as this is the most reliable proof for the presence of live bacteria. Moreover, a successful trial will enable a broader molecular characterization and speciation of isolated colonies. The present guideline gathers and summarizes recommendations, including advantages and limitations of isolation and molecular detection of Bartonella from mammalian and arthropod samples

Comparison of Simultaneous Splenic Sample PCR with Blood Sample PCR for Diagnosis and Treatment of Experimental Ehrlichia canis Infection

This report presents evidence that dogs recover from acute canine monocytic ehrlichiosis (CME) after 16 days of doxycycline treatment (10 mg/kg of body weight every 24 h). Blood PCR was as valuable as splenic aspirate PCR for early diagnosis of acute CME. Splenic aspirate PCR was, however, superior to blood PCR for the evaluation of ehrlichial elimination