Clinical management of treatment failure in patients with oropharyngeal tularemia: A retrospective evaluation

Tularemia is a zoonotic disease caused by Francisella tularensis pathogen. It can be transmitted via wild animals, infected water, and contaminated food. In our study, 13 patients who were admitted to our clinic with a neck mass were retrospectively evaluated for symptoms, findings, diagnosis, applied treatments, and the causes of delay in treatment. The ages of patients diagnosed with tularemia ranged from 19–70 years, with a mean age of 48.5 years. All patients resided in the countryside and all had been repeatedly administered penicillin and macrolide group antibiotics before referral to our clinic. The patients consulted our clinic with a delay of 8 to 30 days (average 11.5 days) after the onset of the first symptom. Six (46.1%) patients had deep lymphadenopathy, while lymphadenopathy in seven (53.8%) was superficial. Suppuration was observed in all adenopathies located superficially to the lymph, while skin fistulization was present in three patients. The diagnosis was made by a serum microagglutination test. Doxycycline, ciprofloxacin, and streptomycin group antibiotics were used in the treatment. No complications due to treatment were observed in the early period. In conclusion; the diagnosis of tularemia is not straightforward at the initial admission of the patients since the symptoms of tonsillopharyngitis, fever, and cervical adenopathy are similar in oropharyngeal tularemia and upper respiratory tract infections. Therefore, the antibiotics administered to the patients are not effective. In endemic regions, tularemia should be considered in the differential diagnosis in patients with tonsillopharyngitis and adenopathy in the neck.

Serological Survey on the Occurrence of Anti-Leptospira spp. Antibodies in Red-Eared Terrapins (Trachemys scripta elegans) Living in a Natural Park of Northern Italy

Turtles are suspected to be involved in the epidemiology of Leptospira; however, data about the dissemination of this zoonotic pathogen among chelonians are scant. In the present study, the serum samples collected from 49 Trachemys scripta elegans living in a natural park of northern Italy were tested by a microagglutination test to measure detectable antibodies against different Leptospira serovars. Three (6.12%) turtles had agglutinins to the serovar Tarassovi, suggesting that they were exposed to the spirochaetes. Currently, it is not clear if Leptospira can cause disease in chelonians or if these animals can serve as reservoirs of leptospirae. Considering that chelonians often share the same environment with other animals and humans, and considering the One Health perspective, investigations to better understand the role of chelonians as a source of Leptospira infection are necessary.

Usefulness of IgM-ELISA test for screening of Leptospirosis in Cuba

Introduction: Leptospirosis is a common cause of acute febrile illness in many tropical regions of the world. Early diagnosis is essential, since untreated cases can progress rapidly and mortality rates are high in severe cases. According to the observations of the Cuban National Reference Laboratory, non-reactive serology’s are prevailing in most suspected cases of human leptospirosis. Objective: to apply the IgM-ELISA test for screening of IgM antibodies using sera from patients with the acute phase of the illness. Material and methods: in the current study, 31 pairs of sera and 140 single sera from 337 suspected patients with leptospirosis were tested by two methods, a commercial IgM-ELISA test for Leptospira and microagglutination test (MAT). Results: IgM-ELISA test results were concordant with MAT results in 90.0% (28/31) of paired sera and 88,6% (124/140) of single sera. The following serogroups: Icterohaemorrhagiae 23,74% (18/76), Pomona 22.3% (17/76), Canicola 13.1% (10/76), and Ballum 5.2% (4/76) were the most frequently found in sera testing positive by IgM-ELISA. Positive IgM-ELISA sera were predominantly those taken from 5th to the 8th day of the acute phase of the illness. Some samples taken from day zero to the 28th day were also positive, suggesting a high sensitivity of this test. Conclusion: IgM-ELISA test is useful for screening of human leptospirosis, particularly if using sera taken from days 5-8 of surveillance, which reduce the under reporting of leptospirosis cases in Cuba.

Behavioural change: a rare presentation of leptospirosis

Neurological manifestations of leptospirosis without severe multiorgan involvement are a rare clinical entity. Despite the increasing prevalence of the disease in many tropical countries, its protean clinical presentations make its timely diagnosis challenging. We report the case of a 44-year-old Filipino man presenting with fever, myalgia, behavioural changes and altered sensorium. Neurological examination did not show any focal neurological deficits or clear signs of meningoencephalitis. Lumbar tap, cranial CT scan and cranial MRI were inconclusive. The diagnosis of leptospirosis with acute encephalitis relied heavily on the patient’s clinical clues, appropriate exposure history and patterns in ancillary laboratory tests. Empiric antibiotic therapy with ceftriaxone was initiated. Seroconversion and fourfold increase in serological antibody titres by leptospirosis microagglutination test later confirmed the diagnosis. The patient was successfully treated, and all neurological complications were reversed.

Use of geoinformation technologies (GIS) in the system of monitoring and prevention of animal leptospirosis of the Cherkas region

The article deals with the application of geographic information technologies (GIS) as a system of monitoring and prevention of animal leptospirosis in Cherkasy region. The analysis of modern programs, where GIS monitoring in veterinary medicine is used, has been carried out. The structure of the database monitoring of animal leptospirosis in Cherkasy region with GIS being used, should include the regional data on: geographic indicators, wildlife habitats and pastures; farm, wild and domestic animals in number; categories of farms; pedigree value; veterinary and other institutions that provide the accounts with their mandatory inclusion in the automatic digital calculating programs (Excel). In 2012 – 2017 the animals in Cherkasy region numbered in 838212 head of cattle, 75383 head of small cattle,1607283 head of pigs, 15195 head of horses. The density of different categories of farms raising these animal species has even distribution; among them 12 districts have dairy farms, 1 district - meat farms, 6 districts - dairy and meat farms. The number of wild animals in Cherkasy region in 2017 was: elk – 81 head; deer – 331 head; roe deer – 6783 head; wild boars – 2380 head, foxes – 2323 head; they were evenly distributed on 1574.9 thousand hectares,and 30 hunting grounds. During the determination of epizootic situation of animal leptospirosis in Cherkasy region in 2012 – 2017, 95106 animals were tested, among them 5078 animals appeared positive for leptospirosis in microagglutination test (MAT), out of which – 4454 head of cattle and pigs - together 87.71%; other species, including wild animals – 624 head (12.29%). The density of leptospirosis cases in the districts among all animal species ranges from 0 to 20%, the highest density is in Cherkasy region; in three areas there was no reporting on any positive cases. Species composition of animals with leptospirosis in Cherkasy region in 2012–2017 was determined by: cattle – 89.17%, small cattle – 0.20, horses – 2.68, pigs – 7.82%; wild goats – 0.02% and wild boars –0.06%; dogs - 0.02%, cats - 0.04%. The etiological profile of animal leptospirosis in Cherkasy region in 2012–2017 consisted mainly of mixed serogroup types - Pomona, Australis, Canicola, Grippotiphosa, Tarassovi, Icnterohaemorrhagiae –32.79%; in the second place - mixed Hebdomadis and Sejroe –14.09%; in the third – Australis –14.17%; separately - Hebdomadis and Sejroe - 11.01 and 11.81%, respectively, Icnterohaemorrhagiae - 8.08%. All others - Pomona Canicola, Grippotiphosa, Tarassovi - from 3 to 0,5%. Adoption of the effective preventative measures of animal leptospirosis in Cherkasy region has become possible on the basis of the complete informative database, awareness about the animal species and their number - among those being registered and tested for leptospirosis, their etiological structure in the above mentioned region with GIS being used.

Diagnosis of canine leptospirosis: evaluation of two PCR assays in comparison with the microagglutination test

ABSTRACT: Canine leptospirosis is definitely diagnosed by demonstrating seroconversion in paired serum samples from the acute and convalescent period by the microagglutination test (MAT). However, the application of a polymerase chain reaction (PCR) assay can provide earlier confirmation of suspected cases. The objective of this study was to evaluate two PCR assays used in diagnosis of human leptospirosis (lipL32 real-time PCR and rrs conventional PCR) in cultured microorganisms and experimentally contaminated samples (whole blood, serum, urine), and investigate their applicability in clinical samples from dogs with presumptive diagnosis of leptospirosis by using the MAT as a reference. The analytical sensitivity of the lipL32 real-time PCR was 1 genome equivalent per reaction, whereas that for the rrs conventional PCR was 10 genome equivalents per reaction. Both assays amplified the pathogenic strains but were negative when evaluating the DNA of other microorganisms that may be present in clinical samples. The lipL32 real-time PCR detected 100 bacteria/mL in whole blood samples, 1000 bacteria/mL in serum samples and 10 bacteria/mL in urine samples, whereas the rrs conventional PCR detected 1000 bacteria/mL in whole blood and serum samples and 100 bacteria/mL in urine samples. Seven out of the 51 samples from dogs with presumptive diagnosis of leptospirosis were considered as confirmed cases. ThelipL32 real-time PCR detected positive results in six of the seven confirmed cases, whereas the rrs conventional PCR detected four. The PCR assays evaluated proved to be useful diagnostic tools in the confirmation of canine leptospirosis when used together with the MAT.

Francisella tularensis in Swedish predators and scavengers

Tularaemia is a zoonotic disease, in Europe caused by Francisella tularensis subsp. holarctica. Many lagomorphs and a variety of small rodents are wildlife species prone to develop clinical disease, while predators and scavengers are relatively resistant and may serve as sentinels. Blood samples from 656 Swedish wild predators and scavengers were serologically investigated using slide agglutination and microagglutination. In the slide agglutination test, 34 seropositive animals were detected, and they were found among all species investigated: brown bear (Ursus arctos), Eurasian lynx (Lynx lynx), raccoon dog (Nyctereutes procyonoides), red fox (Vulpes vulpes), wild boar (Sus scrofa), wolf (Canis lupus) and wolverine (Gulo gulo). Due to haemolysis the microagglutination test was more difficult to read at low titres, and only 12 animals were classified as seropositive. F. tularensis subsp. holarctica was detected by a polymerase chain reaction in lymphatic tissues of the head in one brown bear, one red fox and one wolf. The significance of this finding regarding possible latency of infection is not clear. In conclusion, the results of this study indicate that all predator and scavenger species included in this study may serve as sentinels for tularaemia in Sweden. Their role as reservoirs is unclear.

Laboratory investigation of adult small ruminant Leptospirosis, a neglected infection in Greece: problems and recommendations

Leptospirosis is in Greece a neglected infection. Small ruminants and specifically sheep are accidental hosts of Leptospira spp, but they could also be disseminators of pathogenic serovars. Thus, the objective was to investigate leptospirosis of adult small ruminants coming from areas in Southern Greece, where accidental evidence had showed that leptospirosis could be an important infection for man and animals. For this purpose, blood and kidney samples were collected at slaughter from adult females. Collected samples were examined with a commercial serological screening kit, the microagglutination test ( MAT), histology and PCR. One hundred ten serum and 110 tissue samples were collected. Of the examined serum samples 55 (50%) were suspect for leptospirosis in the screening kit and 28 (25.45%) were MAT positive. Of the tissue samples 38 (34.5%) were PCR positive and 30 (27.2%) showed various degrees of microscopic kidney lesions. The serovars identified by the MAT were Tarassovi (10 animals), Autumnalis (8 animals), Zanoni (4 animals), Hebdomadis and Javanica (2 each), Bratislava and Hardjio prajitno (one each). The conclusion is that small ruminants and specifically sheep (98 animals) are disseminators of pathogenic Leptospira spp. serovars in areas where they predominate and climatic factors favor the survival of the pathogen.

Evaluation of In-House and Commercial Serological Tests for Diagnosis of Human Tularemia

ABSTRACTTularemia is a zoonosis caused by the bacteriumFrancisella tularensis. Its specific diagnosis remains based on serological methods, whileF. tularensisis rarely detected in clinical samples by culture or PCR. The aim of the present study was to evaluate the performance of the Serion enzyme-linked immunosorbent assay (ELISA) classicFrancisella tularensisIgG and IgM tests (Virion/Serion GmbH Institute, Würzburg, Germany) and the VIRapid tularemia immunochromatographic test (ICT) (Vircell, Granada, Spain) compared to that of the in-house microagglutination test (MAT) and indirect immunofluorescence assay (IFA) currently used at the French National Reference Center forFrancisella. We evaluated 256 consecutive sera from 208 patients, including 51 confirmed and 23 probable tularemia cases, and 134 control patients not infected withF. tularensis. The IFA tests displayed 72.5% sensitivity for IgM (cutoff titer ≥80) and 74.5% for IgG (cutoff titer ≥160), and 99.3% specificity for both IgM and IgG. Using cutoffs advocated by the manufacturer, the Serion ELISAs displayed 88.2% sensitivity for IgM and 86.3% for IgG antibodies; specificity was 94.8% for IgM and 95.5% for IgG. Compared to MAT and IFA tests, the Serion ELISAs allowed earlier detection of specific antibodies (1 to 2 weeks versus 2 to 3 weeks after the onset of symptoms). The ICT sensitivity and specificity were 90% and 83.6%, respectively, when considering the cutoff advocated by the manufacturer. In conclusion, the Serion ELISAs are useful as screening tests for tularemia diagnosis, but additional confirmatory tests (such as MAT and IFA) are needed, especially in areas of low endemicity.