Antibodies against Borrelia burgdorferi Sensu Lato in Clinically Healthy and Sick Horses: First Report from the Czech Republic.

Lyme disease, caused by some strains of bacterial spirochetes Borrelia burgdorferi sensu lato (Bbsl), affects humans but also domestic animals including horses. The primary pathogens in horses in Europe are B. afzelii, B. garinii and B. burgdorferi sensu stricto. To our knowledge, there are no data available on the seropositivity of B. burgdorferi s.l. in horses from the Czech Republic. In this country, horses are mainly used for sport, breeding, and recreational riding in areas where vectors of B. burgdorferi s.l. are present, which is why they are frequently at risk of infection. The aim of the study was to detect anti-borrelia IgM and IgG antibodies in clinically healthy and sick horses from the Czech Republic and to evaluate the risk factors of infection. In total, sera of 262 horses (247 clinically healthy horses and 15 horses hospitalized due to symptoms of encephalitis/meningoencephalitis) were examined by an indirect sandwich enzyme-linked immunosorbent assay. Positivity of B. burgdorferi was 27% (66/247) in clinically healthy horses (21% IgM, 7% IgG and 3% IgM + IgG antibodies) and 20% (3/15) in horses with clinical signs (20% IgM, 7% IgG and 7% IgM + IgG). In the clinically healthy horses, positivity statistically differed (p ≤ 0.05) only in Pony and Warmblood breeds, being the most affected at 32% and 30%, respectively, while other characteristics (sex, age, usage and localities) had no effect on positivity. This is the first survey of antibodies to B. burgdorferi s.l. in Czech horses showing that horses are exposed to ticks infected with B. burgdorferi s.l. This should be taken into account when making differential diagnoses in patients with non-specific symptoms to start with adequate therapy.

[Monitoring the activity of Ixodes ricinus ticks and their positivity for selected pathogenic microorganisms in the agglomeration of the city of Brno].

OBJECTIVE: The aim is to present the results of monitoring the activity of Ixodes ricinus ticks and their positivity for dangerous pathogenic microorganisms in the Brno-Pisarky forest park. The monitored pathogens were Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Coxiella burnetii, Erhlichia chaffensis and Francisella tularensis. Additionally, factors potentially influencing tick activity in the location (especially the temperature) were monitored. MATERIALS AND METHODS: In the location, ticks were caught using the flagging method. Positivity of ticks was determined by polymerase chain reaction followed by gel electrophoresis. RESULTS: Between March and November 2021, 563 ticks were collected in the location, the vast majority being nymphs (85 %). The positivity rates were 5.6 % for the pathogen Bbsl was and 2.4 % for A. phagocytophilum. C. burnetii, E. chaffensis and F. tularensis were not confirmed in the location. CONCLUSION: The location was found to be infested with the pathogens Bbsl and A. phagocytophilum. However, the positivity in Brno-Pisarky is lower compared to other Brno areas.

Leptospira interrogans Sensu Lato in Wild Small Mammals in Three Moravian Localities of the Czech Republic.

Leptospirosis is a widespread zoonosis, affecting humans, domestic animals and wildlife, with small mammals as a reservoir of this infection. In recent years, this disease has been re-emerging and affects approximately 1 million people all over the world each year. Due to this disease having a significant health impact, it is important to identify the source and method of infection. The risk of Leptospira sp. infection is higher mainly in the cities of developed and industrialised countries. The aim of the study was the detection of antibodies against Leptospira sp. in some wild small mammals captured in the Czech Republic. In total, samples of 855 animals captured in three locations of Moravia during a six-year study (2010-2015) were examined by a microscopic agglutination test, using eight serovars of Leptospira interrogans sensu lato, representing serogroups Grippotyphosa, Icterohaemorrhagiae, Australis, Canicola, Sejroe, Javanica, Pomona and Pyrogenes, as antigens. Antibodies to Leptospira sp. were detected in 6.1% (52/855) of animals, with a prevalence of 6.4% (51/801) and 1.9% (1/54) in rodents and insectivores, respectively. The only statistically significant difference (p ≤ 0.05) was in prevalence between individual species (0-33%), while there were no differences in sex (6.7% in females and 5.1% in males), locality (1.8-8%) and year of trapping (0-8.4%). Only two serovars, L. interrogans serovar Pomona and L. interrogans serovar Grippotyphosa, were detected in 5.5% and 0.5% of animals, respectively. The prevailing serovar of pathogenic L. interrogans s.l. can be identified in a number of infected people in the Czech Republic. The composition of vaccines should be based on the current occurrence of Leptospira serovars in the actual territory. For this reason, the occurrence of Leptospira and its serovars should therefore be regularly monitored.

Wild Small Mammals and Ticks in Zoos-Reservoir of Agents with Zoonotic Potential?

Wild small mammals and ticks play an important role in maintaining and spreading zoonoses in nature, as well as in captive animals. The aim of this study was to monitor selected agents with zoonotic potential in their reservoirs and vectors in a zoo, and to draw attention to the risk of possible contact with these pathogens. In total, 117 wild small mammals (rodents) and 166 ticks were collected in the area of Brno Zoo. Antibodies to the bacteria Coxiella burnetii, Francisella tularensis, and Borrelia burgdorferi s.l. were detected by a modified enzyme-linked immunosorbent assay in 19% (19/99), 4% (4/99), and 15% (15/99) of rodents, respectively. Antibodies to Leptospira spp. bacteria were detected by the microscopic agglutination test in 6% (4/63) of rodents. Coinfection (antibodies to more than two agents) were proved in 14.5% (15/97) of animals. The prevalence of C. burnetii statistically differed according to the years of trapping (p = 0.0241). The DNAs of B. burgdorferi s.l., Rickettsia sp., and Anaplasma phagocytophilum were detected by PCR in 16%, 6%, and 1% of ticks, respectively, without coinfection and without effect of life stage and sex of ticks on positivity. Sequencing showed homology with R. helvetica and A. phagocytophilum in four and one positive samples, respectively. The results of our study show that wild small mammals and ticks in a zoo could serve as reservoirs and vectors of infectious agents with zoonotic potential and thus present a risk of infection to zoo animals and also to keepers and visitors to a zoo.

Antibodies Related to Borrelia burgdorferi sensu lato, Coxiella burnetii, and Francisella tularensis Detected in Serum and Heart Rinses of Wild Small Mammals in the Czech Republic.

Wild small mammals are the most common reservoirs of pathogenic microorganisms that can cause zoonotic diseases. The aim of the study was to detect antibodies related to Borrelia burgdorferi sensu lato, Coxiella burnetii, and Francisella tularensis in wild small mammals from the Czech Republic. In total, sera or heart rinses of 211 wild small mammals (168 Apodemus flavicollis, 28 Myodes glareolus, 9 A. sylvaticus, and 6 Sorex araneus) were examined by modified enzyme-linked immunosorbent assay. Antibodies related to B.burgdorferi s.l., C. burnetii, and F. tularensis were detected in 15%, 19%, and 20% of animals, respectively. The prevalence of B. burgdorferi and F. tularensis statistically differed in localities and F. tularensis also differed in sex. Antibodies against 2-3 pathogens were found in 17% of animals with a higher prevalence in M. glareolus. This study brings new data about the prevalence of the above-mentioned pathogens.

Seroprevalence of Antibodies against Borrelia burgdorferi s. l. and Leptospira interrogans s. l. in Cats in district of Brno and its environs, the Czech Republic.

OBJECTIVES: The aim of this study is to evaluate the seroprevalence of antibodies of Borrelia burgdorferi sensu lato (Bbsl) and Leptospira interrogans sensu lato (Lisl) and their possible concurrence in domestic cats living in variable conditions in South Moravia in the district of Brno and its environs. Additional objectives were to discover possible differences in seroprevalence between groups of cats living in different living conditions, and to determine the spectrum of Leptospira serogroups in cats in the same places. MATERIAL AND METHODS: A total of 360 blood sera from domestic cats of 3 different sets were collected during the period 2013-2015. All samples were examined using ELISA for the detection of IgM and IgG antibodies against Bbsl, and the microscopic agglutination test (MAT) for the detection of antibodies against 8 serogroups of Lisl. RESULTS: The ELISA method determined 15.8%, 4.8% and 10.3% IgM anti-Borrelia antibodies in the patient group, shelter cats and street cats, respectively. IgG anti-Borrelia antibodies were found in 6.2%, 9.5%, 5.2%, respectively. Antibodies specific for 5 Leptospira serogroups were detected by the use of MAT in 8.8%, 9.5% and 10.3% of cats from the investigated groups. The total positivity of all examined cats for anti-Borrelia antibodies was 18.0% and for anti-Leptospira - 9.2%. CONCLUSIONS: Cats can be infected with both Bbsl and Lisl. The obtained results are exclusive to the city of Brno and its environs, and are comparable to the limited previous studies. There is a need for further studies of clinical signs of both infections and the possible transmission of Leptospira by ticks.

Coxiella burnetii and Francisella tularensis in wild small mammals from the Czech Republic.

Wild rodents are an important source of the tick-borne pathogens Coxiella burnetii and Francisella tularensis. The aim of our study was to assess the prevalence of antibodies and possible coexistence of these pathogens in wild small mammals from three localities in the Czech Republic. A total of 614 wild small mammals (324 Apodemus flavicollis, 145 Myodes glareolus, 50 Sorex araneus, 48 A. sylvaticus, 40 A. agrarius, six Microtus arvalis and one Talpa europaea) were trapped between 2012 and 2015. Their sera or heart extracts were examined by modified indirect enzyme-linked immunosorbent assay, with the detection of antibodies against C. burnetii and F. tularensis in 12 % and 7 % of animals, respectively; coinfection was identified in 4.4 % of animals. The prevalence of C. burnetii and F. tularensis antibodies statistically differed according to animal species and sex (p < 0.05); the seroprevalence of C. burnetii (p < 0.05) also differed in the sampling period. The highest prevalence of antibodies against C. burnetii and F. tularensis was detected in the case of M. glareolus (24 % and 14 %, respectively).

Molecular Epidemiology of Hantaviruses in the Czech Republic.

During 2008-2018, we collected samples from rodents and patients throughout the Czech Republic and characterized hantavirus isolates. We detected Dobrava-Belgrade and Puumala orthohantaviruses in patients and Dobrava-Belgrade, Tula, and Seewis orthohantaviruses in rodents. Increased knowledge of eco-epidemiology of hantaviruses will improve awareness among physicians and better outcomes of patients.

Body Composition Changes During a 24-h Winter Mountain Running Race Under Extremely Cold Conditions.

Background: To date, no study has focused on body composition characteristics and on parameters associated with skeletal muscle damage and renal function in runners participating in a 24-h winter race held under extremely cold environmental conditions (average temperature of -14.3°C). Methods: Anthropometric characteristics, plasma urea (PU), plasma creatinine (Pcr), creatine kinase (CK), plasma volume (PV) and total body water (TBW) were assessed pre- and post-race in 20 finishers (14 men and 6 women). Results: In male runners, body mass (BM) (p = 0.003) and body fat (BF) (p = 0.001) decreased [-1.1 kg (-1.4%) and -1.1 kg (-13.4%), respectively]; skeletal muscle mass (SM) and TBW remained stable (p > 0.05). In female runners, BF decreased (p = 0.036) [-1.3 kg (-7.8%)] while BM, SM and TBW remained stable (p > 0.05). The change (Δ) in BM was not related to Δ BF; however, Δ BM was related to Δ SM [r = 0.58, p = 0.007] and Δ TBW (r = 0.59, p = 0.007). Δ SM correlated with Δ TBW (r = 0.51, p = 0.021). Moreover, Δ BF was negatively associated with Δ SM (r = -0.65, p = 0.002). PV (p < 0.001), CK (p < 0.001), Pcr (p = 0.004) and PU (p < 0.001) increased and creatinine clearance (CrCl) decreased (p = 0.002). The decrease in BM was negatively related to the increase in CK (r = -0.71, p < 0.001). Δ Pcr was positively related to Δ PU (r = 0.64, p = 0.002). The decrease in CrCl was negatively associated with the increase in both PU (r = -0.72, p < 0.001) and CK (r = -0.48, p = 0.032). Conclusion: The 24-h running race under extremely cold conditions led to a significant BF decrease, whereas SM and TBW remained stable in both males and females. Nevertheless, the increase in CK, Pcr and PU was related to the damage of SM with transient impaired renal function.

Detection of antibodies to Borrelia burgdorferi s.l. in wild small mammals and sensitivity of PCR and cultivation.

The aim of this study was to determine the seroprevalence of antibodies to B. burgdorferi s.l. in wild small mammals in the Czech Republic and compare sensitivity of PCR and cultivaton. Wild small mammals (n = 691) were trapped in years 2010-2014 in three localities of the Czech Republic. Heart rinses (n = 340) and sera (n = 351) were examined by modified indirect ELISA. Seventy animals were randomly selected for comparison of results of cultivation and PCR. Mean annual antiborelian positivity was 12% with statistical difference (p < 0.05) between Bank Vole (Clethrionomys glareolus) and other six animal species, while there was no statistical difference (p > 0.05) between rodentia and insectivora, gender and localities. The cultivation revealed one positive sample (1.4%), negative in both PCR and ELISA. Method PCR revealed seven positive samples (10%); two of them were simultaneously dubious in ELISA. Eleven animals, negative in cultivation and PCR, had antibodies in ELISA. Method of PCR compared to cultivation seems to be more sensitive for detection of Borrelia.

Pilot study of Ixodes ricinus ticks preference for human ABO blood groups using a simple in vitro method.

INTRODUCTION AND OBJECTIVES: The existence of a blood group preference for ticks is a problem widely discussed among the lay public but often neglected by the scientific community. The Ixodes ricinus tick transmits serious zoonotic diseases such as Lyme borreliosis, tick encephalitis, or anaplasmosis. The preventive strategies include vaccination (if available) and individual measures including the use of repellents and avoidance of risk areas. MATERIAL AND METHODS: Since this topic is relatively neglected in the scientific literature, a simple in vitro method was used. Ticks used in this study were collected in the suburban region of Ruda in Brno, Czech Republic. One hundred active nymphs of the collected ticks were tested for preferences for blood groups, using Petri dishes and blood samples from volunteers. To demonstrate the threat of ticks and the diseases they transmit, the positivity of one of the most abundant zoonosis, Lyme borreliosis, was tested using dark-field microscopy. RESULTS: The results obtained showed that the examined ticks were attracted most by blood group A, whereas the least preferred was group B, which was proved statistically (p <0.05). The mean positivity of collected ticks for the presence of spirochaetes was 9.35%. CONCLUSIONS: The results indicate that the host selection of ticks may be influenced by the physiological or biochemical profile of an individual, such as their blood group. This means that a blood group of an individual can be one of the factors that increase the risk of tick bite and the transmission of dangerous diseases and thus must not be underestimated.

The Effect of a 100-km Ultra-Marathon under Freezing Conditions on Selected Immunological and Hematological Parameters.

Although moderate exercise is beneficial for the human body and its immune system, exhaustive ultra-endurance performance in cold conditions might be harmful. The aim of this study was to examine the effect of a 100-km ultra-marathon under cold conditions (temperatures from -1°C to +1°C) on selected immunological, biochemical and hematological parameters. Participants were 15 runners (12 men and three women, age 40.3 ± 9.7 years, body mass 67.3 ± 9.0 kg and body height 1.74 ± 0.10 m, mean ± standard deviation). Leukocytes increased (p < 0.01) and, particularly, the number of leucocytes doubled in seven out of 15 athletes. Immature neutrophils, mature neutrophils and monocytes increased (p < 0.02), whereas lymphocytes and eosinophils did not change. IgG increased (p < 0.02), but IgA and IgM remained unchanged. Platelets increased (p < 0.01), whereas red blood cells, hematocrit and hemoglobin did not change. lactate dehydrogenase (LDH) and creatine kinase (CK) increased (p < 0.01), but alanine aminotransferase (ALT) did not change. There was an association between the markers of the acute inflammation of the organism (i.e., neutrophils, immature neutrophils, platelets, and monocytes) and the markers of muscle damage (i.e., CK, platelets, and LDH). There were no relationships among all the markers in relation to upper respiratory tract infections and liver damage. The highest change was noted in the increase of the number of immature neutrophils (1,019.2%) and CK levels (1,077.6%). In summary, this is the first study investigating immunological, hematological and biochemical parameters and showing that running a 100-km ultra-marathon under cold conditions leads to changes in several immunological, biochemical and hematological parameters indicating a severe stress on the body associated with increasing susceptibility to the development of infections.

Description of Three Female 24-h Ultra-Endurance Race Winners in Various Weather Conditions and Disciplines.

A The incidence of exercise-associated hyponatremia (EAH) is higher in women than in men. We present three cases of a very mild post-race EAH in female winners of three 24-h ultra races in various weather conditions and disciplines with post-race plasma sodium [Na⁺] levels of 134 mM (Case 1), 133 mM (Case 2) and 134 mM (Case 3). Moreover, Case 1 and Case 2 showed elevated creatine kinase concentrations of >10,000 U/l with an absence of renal function abnormality. The common characteristics were female sex, veteran recreational category, long race experience in the particular sports discipline, excellent race performance, similar total weekly training hours and the presence of luteal phase of the menstrual cycle during the race. Hematocrit and hemoglobin decreased and post-race K⁺/Na⁺ ratio in urine increased in all three cases. In addition, an increased body mass and a decreased urine specific gravity and urine osmolality suggested over-drinking in Case 1. A decrease in the glomerular filtration rate and creatine clearance accompanied by an increase in urine [Na⁺] may contribute to fluid overload in Cases 2 and 3. Furthermore, urine osmolality reached a level indicating antidiuretic hormone secretion in all the present cases. Therefore, we recommend that race medical personnel should not forget to look for EAH even in fast and experienced female athletes and during races in different environmental conditions.

Students' attitudes to tick risks.

INTRODUCTION AND OBJECTIVES: The ever-increasing number of patients with tick-borne diseases resulted in the presented study investigating the awareness, attitudes and knowledge among students about the threats arising from tick bites and preventive anti-tick practices. MATERIALS AND METHOD: Questionnaires concerning these issues were distributed amongst Czech and Polish university students of science. Responses were analyzed by nationality and by gender. RESULTS: Nearly all respondents were aware of the risks arising from ticks and could name at least one disease transmitted by ticks. The Czech students felt more threatened by tick-borne diseases, had more frequently suffered from Lyme borreliosis and were vaccinated against tick-borne encephalitis more often than the Polish students. A large number of the participants applied preventive measures against ticks in order to protect themselves. The Czech students believed in the effectiveness of repellents statistically more often than the Polish students, while effectiveness is the main criterion for selection of the right repellent in both groups. CONCLUSION: Differences in preferences between the two nations appeared in many areas, e.g. the Czechs felt more threatened by all kind of risks and suffered from Lyme disease more frequently. Gaps can still be found in both the knowledge and behaviour among the respondents. It can be expected that the general public knowledge of this issue is rather limited in comparison with the students participating in the study, who are systematically educated in the field.

Toxoplasma gondii and Neospora caninum in wild small mammals: Seroprevalence, DNA detection and genotyping.

Generally, rodents and other small mammals are considered as one of the sources of Toxoplasma gondii or Neospora caninum infection for cats and dogs as the definitive hosts of these two parasites, respectively. The aim of the study was to find out the prevalence of these two parasites in wild small mammals from the Czech Republic and to characterize T. gondii isolates by methods of molecular biology. A total of 621 wild small mammals were caught in the Czech Republic during years 2002-2014. Antibodies to T. gondii were detected by latex agglutination test in six (2.5%) of 240 small mammals (in two A. agrarius and four A. flavicollis). Antibodies to N. caninum were detected by commercially available competitive-inhibition enzyme-linked immunosorbent assay in one A. flavicolis (0.4%). Three of 427 (0.7%) liver samples were positive for T. gondii by PCR while negative for N. caninum. All embryo samples (n=102) were negative for both T. gondii and N. caninum. The three liver samples positive for T. gondii DNA (two from A. flavicollis and one from A. sylvaticus) were genotyped by 15 microsatellite markers and characterized as type II. To our knowledge, this is the first information about genetic characterization of T. gondii isolates in small mammals from Europe and the first detection of N. caninum antibodies in wild rodents from the Czech Republic.

DNA-based identification and OspC serotyping in cultures of Borrelia burgdorferi s.l. isolated from ticks collected in the Moravia (Czech Republic).

Two different genetic loci, flaB and ospC, were employed to assign genospecies and OspC phylogenetic type to 18 strains isolated from ticks collected in Pisárky, a suburban park in the city of Brno, Czech Republic. The RFLP analysis revealed three different genospecies (B. afzelii, B. garinii, and B. valaisiana). Three samples from the collection contained more than one genospecies. In the other 15 strains, nucleotide sequences of flaB and ospC were determined. The following phylogenetic analysis assigned 12 isolates to genospecies B. garinii and three to B. afzelii. These isolates were further subdivided into seven distinct ospC groups. The most related OspC types were G2, G4, and G5 (B. garinii) and A3 and A8 (B. afzelii).

Identification of Borrelia burgdorferi genospecies isolated from Ixodes ricinus ticks in the South Moravian region of the Czech Republic.

INTRODUCTION: During 2008-2012, a total of 466 ticks Ixodes ricinus removed from humans were collected and tested for the presence of Borrelia burgdorferi sensu lato (Bbsl). Ticks were collected in all districts of the South Moravian region of the Czech Republic (CZ). OBJECTIVE: The aim of this study was to determine the infestation of Bbsl in ticks Ixodes ricinus and the identification of genospecies of Bbsl group by DNA sequencing. MATERIAL AND METHODS: DNA isolation from homogenates was performed by UltraClean BloodSpin DNA kit (MoBio) and by automated instrument Prepito (Perkin-Elmer). Detection of spirochetes was carried out by RealTime PCR kit EliGene Borrelia LC (Elisabeth Pharmacon). Finally, all the positive samples were sequenced on an ABI 3130 Genetic Analyzer (Life Technologies) and identified in the BLAST (NCBI) database. RESULTS: A total positivity of the samples was 26%. For the first time in the Czech Republic, 5 of the isolated strains were genotyped as Borrelia spielmanii (7.1%). Other representatives of Bbsl were also observed: B. afzelii (70.0%), B. garinii (10.0%), B. valaisiana (8.6%), and B. burgdorferi s. s. (4.3%). CONCLUSION: A general view of the spreading of Bbsl in the South Moravian region was demonstrated. The most interesting result of the study is the finding of B. spielmanii for the first time in this region.

Rhabdomyolysis and exercise-associated hyponatremia in ultra-bikers and ultra-runners.

BACKGROUND: Exercise-associated hyponatremia (EAH), rhabdomyolysis and renal failure appear to be a unique problem in ultra-endurance racers. METHODS: We investigated the combined occurrence of EAH and rhabdomyolysis in seven different ultra-endurance races and disciplines (i.e. multi-stage mountain biking, 24-h mountain biking, 24-h ultra-running and 100-km ultra-running). RESULTS: Two (15.4%) ultra-runners (man and woman) from hyponatremic ultra-athletes (n = 13) and four (4%) ultra-runners (four men) from the normonatremic group (n = 100) showed rhabdomyolysis following elevated blood creatine kinase (CK) levels > 10,000 U/L without the development of renal failure and the necessity of a medical treatment. Post-race creatine kinase, plasma and urine creatinine significantly increased, while plasma [Na(+)] and creatine clearance decreased in hyponatremic and normonatremic athletes, respectively. The percentage increase of CK was higher in the hyponatremic compared to the normonatremic group (P < 0.05). Post-race CK levels were higher in ultra-runners compared to mountain bikers (P < 0.01), in faster normonatremic (P < 0.05) and older and more experienced hyponatremic ultra-athletes (P < 0.05). In all finishers, pre-race plasma [K(+)] was related to post-race CK (P < 0.05). CONCLUSIONS: Hyponatremic ultra-athletes tended to develop exercise-induced rhabdomyolysis more frequently than normonatremic ultra-athletes. Ultra-runners tended to develop rhabdomyolysis more frequently than mountain bikers. We found no association between post-race plasma [Na(+)] and CK concentration in both hypo- and normonatremic ultra-athletes.

Changes in foot volume, body composition, and hydration status in male and female 24-hour ultra-mountain bikers.

BACKGROUND: The effects of running and cycling on changes in hydration status and body composition during a 24-hour race have been described previously, but data for 24-hour ultra-mountain bikers are missing. The present study investigated changes in foot volume, body composition, and hydration status in male and female 24-hour ultra-mountain bikers. METHODS: We compared in 49 (37 men and 12 women) 24-hour ultra-mountain bikers (ultra-MTBers) changes (Δ) in body mass (BM). Fat mass (FM), percent body fat (%BF) and skeletal muscle mass (SM) were estimated using anthropometric methods. Changes in total body water (TBW), extracellular fluid (ECF) and intracellular fluid (ICF) were determined using bioelectrical impedance and changes in foot volume using plethysmography. Haematocrit, plasma [Na+], plasma urea, plasma osmolality, urine urea, urine specific gravity and urine osmolality were measured in a subgroup of 25 ultra-MTBers (16 men and 9 women). RESULTS: In male 24-hour ultra-MTBers, BM (P < 0.001), FM (P < 0.001), %BF (P < 0.001) and ECF (P < 0.05) decreased whereas SM and TBW did not change (P > 0.05). A significant correlation was found between post-race BM and post-race FM (r = 0.63, P < 0.001). In female ultra-MTBers, BM (P < 0.05), %BF (P < 0.05) and FM (P < 0.001) decreased, whereas SM, ECF and TBW remained stable (P > 0.05). Absolute ranking in the race was related to Δ%BM (P < 0.001) and Δ%FM in men (P < 0.001) and to Δ%BM (P < 0.05) in women. In male ultra-MTBers, increased post-race plasma urea (P < 0.001) was negatively related to absolute ranking in the race, Δ%BM, post-race FM and Δ%ECF (P < 0.05). Foot volume remained stable in both sexes (P > 0.05). CONCLUSIONS: Male and female 24-hour ultra-MTBers experienced a significant loss in BM and FM, whereas SM remained stable. Body weight changes and increases in plasma urea do not reflect a change in body hydration status. No oedema of the lower limbs occurred.