Characterizing Lomerizine metabolites in camel urine: High-resolution mass spectrometry method development and validation for enhanced doping control.

RATIONALE: Lomerizine (LMZ) is an antimigraine drug that works as a calcium channel blocker and has selective effects on the central nervous system. It is metabolized into trimetazidine (TMZ), which is a prohibited substance owing to its performance-enhancing effects in both human and animal sports. Effective doping control measures are imperative to distinguish the source of TMZ in samples to ensure integrity and fairness of the sport, therefore a comprehensive analysis of LMZ metabolites is essential to identify potential biomarkers in camel urine for effective doping control. METHODS: Camel urine samples were collected from four healthy animals following a single oral administration of LMZ at a dosage of 1 mg/kg body weight. In vitro studies were conducted using homogenized camel liver samples. Lomerizine and its metabolites were extracted using solid-phase extraction and analyzed with a Thermo Fisher Orbitrap Exploris liquid chromatography mass spectrometry system. The acquired data was processed with the Compound Discoverer software. RESULTS: The study conducted a comprehensive analysis of LMZ metabolites in camels and identified 10 phase I and one phase II metabolites. The primary pathway for the formation of phase I metabolites was de-alkylation, while phase II metabolite was formed through alkylation of the parent drug. The study provided valuable insights into the unique metabolic pathways of LMZ in camels under specific experimental conditions. CONCLUSION: The developed method enables the detection and characterization of LMZ and its metabolites in camels. The identified metabolites has the potential to act as marker metabolites for the distinctive detection of LMZ in camel urine to ensure efficient analytical strategies for routine doping control applications.

Comprehensive metabolite profiling of trimetazidine in camels using high-resolution accurate mass spectrometry: Implications for doping control.

RATIONALE: Trimetazidine and its metabolites are prohibited substances in sports. With a growing number of adverse findings in human athletes, it is crucial to develop doping control strategies that include screening for trimetazidine in animal sports. This study aims to detect and characterize trimetazidine and its metabolites for doping control in camel racing. METHODS: Camel urine and plasma samples were collected from four healthy animals following a single oral dose of trimetazidine. In vitro investigations were conducted using camel liver samples. Liquid-liquid extraction and solid-phase extraction techniques were employed for the extraction of trimetazidine metabolites from plasma and urine matrices. The metabolites were analyzed using a Thermo Orbitrap Exploris LC-MS system with optimized settings to achieve maximum sensitivity and accurate mass measurements. RESULTS: Comprehensive metabolite profiling of trimetazidine in camels revealed the identification of seven phase I and five phase II metabolites. Phase I metabolites were primarily formed through dealkylation, while phase II metabolites were dominated by glucuronide conjugation of demethylated trimetazidine. The findings provided insights into the distinct metabolic pathways and biotransformation patterns of trimetazidine in camels under the experimental conditions. CONCLUSION: The developed method enables detection and characterization of trimetazidine and its metabolites in camels. The identified metabolites have the potential to serve as marker metabolites for trimetazidine abuse in camel racing. This study provides valuable insights into the metabolism of trimetazidine in camels.

Serodiagnosis of aspergillosis in falcons (Falco spp.) by an Afmp1p-based enzyme-linked immunosorbent assay.

Aspergillosis in falcons may be associated with high mortality and difficulties in clinical and laboratory diagnosis. We previously cloned an immunogenic protein, Afmp1p, in Aspergillus fumigatus and showed that anti-Afmp1p antibodies were present in human patients with A. fumigatus infections. In this study, we hypothesise that a similar Afmp1p-based enzyme-linked immunosorbent assay (ELISA) could be applied to serodiagnose falcon aspergillosis. A specific polyclonal antibody was first generated to detect falcon serum IgY. Horseradish peroxidase-conjugate of this antibody was then used to measure anti-Afmp1p antibodies in sera collected from falcons experimentally infected with A. fumigatus, and the performance of the Afmp1p-based ELISA was evaluated using sera from healthy falcons and falcons with documented A. fumigatus infections. All four experimentally infected falcons developed culture- and histology-proven invasive aspergillosis. Anti-Afmp1p antibodies were detected in their sera. For the Afmp1p-based ELISA, the mean ± SD OD450 nm using sera from 129 healthy falcons was 0.186 ± 0.073. Receiver operating characteristics curve analysis showed an absorbance cut-off value of 0.407. One negative serum gave an absorbance outside the normal range, giving a specificity of 99.2%. For the 12 sera from falcons with confirmed aspergillosis, nine gave absorbance values ≥ cut-off, giving a sensitivity of 75%. The Afmp1p-based ELISA is useful for serodiagnosis of falcons with aspergillosis.

Digesta kinetics in gazelles in comparison to other ruminants: Evidence for taxon-specific rumen fluid throughput to adjust digesta washing to the natural diet.

Digesta flow plays an important role in ruminant digestive physiology. We measured the mean retention time (MRT) of a solute and a particle marker in the gastrointestinal tract (GIT) and the reticulorumen (RR) of five gazelles and one dikdik species. Species-specific differences were independent from body mass (BM) or food intake. Comparative evaluations (including up to 31 other ruminant species) indicate that MRT GIT relate positively to BM, and are less related to feeding type (the percentage of grass in the natural diet, %grass) than MRT RR. The MRTparticleRR is related to BM and (as a trend) %grass, matching a higher RR capacity with increasing BM in grazers compared to browsers. MRTsoluteRR is neither linked to BM nor to %grass but shows a consistent phylogenetic signal. Selectivity factors (SF; MRTparticle/MRTsolute, proxies for the degree of digesta washing) are positively related to %grass, with a threshold effect, where species with >20% grass have higher SF. These findings suggest that in different ruminant taxa, morphophysiological adaptations controlling MRTsoluteRR evolved to achieve a similar SF RR in relation to a %grass threshold. A high SF could facilitate an increased microbial yield from the forestomach. Reasons for variation in SF above the %grass threshold might represent important drivers of ruminant diversification and await closer investigation.

Energy requirements and metabolism of the Phillip's dikdik (Madoqua saltiana phillipsi).

Basal metabolic rates in mammals are mainly determined by body mass, but also by ecological factors. Some mammalian species inhabiting hot, dry environments were found to have lower metabolic rates compared to temperate species. We studied energy metabolism in Phillip's dikdik (Madoqua saltiana phillipsi), a small antelope inhabiting xeric shrubland habitats in the Eastern 'horn' of Africa, and compared results to literature data. We measured body mass (BM) changes and digestibility in 12 adults kept on different food intake levels to determine, by extrapolation to zero BM change, maintenance energy requirements (MEm) for metabolizable energy (ME). The MEm averaged at 404±20kJMEkgBM(-0.75)d(-1). In addition we conducted 24h-chamber respirometry with seven fed (non-fasted) individuals. Their mean metabolic rate as calculated from oxygen consumption was 403±51kJkgBM(-0.75)d(-1), corroborating the results of the feeding experiments. Selecting the 20 lowest values of the respiration measurement period to estimate resting metabolic rate (RMR) resulted in a mean RMR of 244±39kJkgBM(-0.75)d(-1), which was not significantly lower than the expected basal metabolic rate of 293kJkgBM(-0.75)d(-1). Therefore, resting metabolism was similar to the expected average basal metabolism of a mammal of this size, which suggests a comparatively low metabolic rate in dikdiks. Compared to literature data Phillip's dikdiks have a MEm similar to measurements reported for small domestic ruminants, but considerably lower than those reported for other wild ruminant species inhabiting temperate and cold climates.

Blood values of captive beira antelope (Dorcatragus megalotis) prior to and during an outbreak of fibrinous pleuropneumonia syndrome (FPPS).

Currently the only captive population of beira antelope (Dorcatragus megalotis) is held at the Al Wabra Wildlife Preservation, Qatar. An outbreak of a severe respiratory disease--fibrinous pleuropneumonia syndrome, most likely caused by Mycoplasma ovipneumoniae--led to a marked population decline. Reactive systemic inflammatory (AA) amyloidosis was noted as a chronic manifestation of the disease. Blood samples had been collected for biochemistry and hematology baseline values prior to the outbreak. Population-level changes were analyzed before and during the course of the outbreak in selected blood parameters (white blood cells [WBC], blood urea nitrogen [BUN], and creatinine). The annual population WBC increased and decreased concurrently with the population size, with a significant correlation between the two measures (R = 0.92; P = 0.001). Both BUN and creatinine values were higher during the outbreak. These values peaked at the same time as mortality, which was 1 yr after the WBC peak. These changes were interpreted as the transition from an acute disease with a primary respiratory manifestation into a chronic condition where renal amyloidosis led to chronic renal failure and death. Also, elevated liver values in diseased animals were attributed to amyloidosis. Parallels to a literature report on a lung disease complex caused by M. ovipneumoniae in bighorn sheep (Ovis canadensis) were found. Trends in population-level blood values of the beira antelopes implicate amyloidosis as a significant, long-term consequence of the putative Mycoplasma infection.

On Serratspiculum (Nematoda; Dicheilonematidae) species occurring in hunting falcons in the United Arab Emirates in respect with their origin.

During two falcon seasons (2020/21 and 2021/22) we investigated Serratospiculum samples from 112 falcons and examined a total of 760 nematodes. Of the 112 falcons, there were 62 Saker (Falco cherrug), 15 Peregrine (Falco peregrinus), 11 Gyr (Falco rusticolus), 7 Lanner falcons (Falco biarmicus) and 17 hybrid falcons. In 47 samples the origin of the birds was not mentioned, 49 were from Mongolia, 6 from the UAE, 3 from the USA, 2 from Russia, 2 from the UK and one each from Kazakhstan, Uzbekistan and Germany. Three different Serratospiculum species were identified: S. seurati from 100 falcons (62 Saker, 11 Gyr, 10 Peregrine, 5 Lanner, 12 hybrid falcons, S. tendo from 10 falcons (4 Peregrine, 2 Lanner, 1 Gyr, three hybrid falcons) and S. guttatum from two falcons (one Peregrine and one hybrid falcon). The main morphological features of the three parasite species were described briefly and hosts and countries where Serratospiculum spp. were found were listed according to references.

Retention of solutes and different-sized particles in the digestive tract of the ostrich (Struthio camelus massaicus), and a comparison with mammals and reptiles.

Ostriches (Struthio camelus) achieve digesta retention times, digesta particle size reduction and digestibilities equal to similar-sized herbivorous mammals, in contrast to some other avian herbivores. The sequence of digestive processes in their gastrointestinal tract, however, is still unexplored. Using two groups of four ostriches (mean body mass 75.1 ± 17.3 kg) kept on fresh alfalfa, we tested the effect of two intake levels (17 and 42 g dry matter kg(-0.75)d(-1)) on the mean retention time (MRT) of a solute and three different-sized (2, 10, 20 mm) particle markers, mean faecal particle size (MPS), and digestibility. Intake level did not affect MRT, but MPS (0.74 vs. 1.52 mm) and dry matter digestibility (81 vs. 78%). The solute marker (MRT 22-26 h) was excreted faster than the particle markers; there was no difference in the MRT of 10 and 20 mm particles (MRT 28-32 h), but 2mm particles were retained longer (MRT 39-40 h). Because the solute marker was not selectively retained, and wet-sieving of gut contents of slaughtered animals did not indicate smaller particles in the caeca, the long MRT of small particles is interpreted as intermittent excretion from the gizzard, potentially due to entrapment in small grit. The marker excretion pattern also showed intermittent peaks for all markers in five of the animals, which indicates non-continuous outflow from the gizzard. When adding our data to literature data on avian herbivores, a dichotomy is evident, with ostrich and hoatzin (Opisthocomus hoazin) displaying long MRTs, high digestibilities, and gut capacities similar to mammalian herbivores, and other avian herbivores such as grouse, geese or emus with shorter MRTs, lower fibre digestibilities and lower gut capacities. In the available data for all avian herbivores where food intake and MRTs were measured, this dichotomy and food intake level, but not body mass, was related to MRT, adding to the evidence that body mass itself may not be sole major determinant of digestive physiology. The most striking difference between mammalian and avian herbivores from the literature is the fundamentally lower methane production measured in the very few studies in birds including ostriches, which appears to be at the level of reptiles, in spite of general food intake levels of a magnitude as in mammals. Further studies in ostriches and other avian herbivores are required to understand the differences in digestive mechanisms between avian and mammalian herbivores.

Malignant lymphoma of T-cell origin in a Humboldt penguin (Spheniscus humboldti) and a pink-backed pelican (Pelecanus rufescens).

Multicentric T-cell lymphomas were diagnosed in two birds from separate zoological collections: one in a 27-year-old female Humboldt penguin (Spheniscus humboldti) and the second in an adult pink-backed pelican (Pelecanus rufescens). The main clinical sign in the penguin was dysphagia caused by lymphoma formation in the esophagus. Besides the esophageal lymphoma, neoplastic lymphoid cells were observed in the adrenal glands, liver, kidneys, lung, proventriculus, and gizzard. The pelican was found dead without a clinical history. Neoplastic lymphoid cells were observed in the kidneys, liver, pancreas, spleen, ventriculus, and small intestine. Neoplastic cells of the penguin as well as of the pelican were immunoreactive to CD3 antigen, suggesting the lymphomas were of T-cell origin. In both cases, test results were negative for Marek's disease virus, avian leukosis virus, and reticuloendotheliosis virus. In the pelican, a skin melanoma was diagnosed on the left throat pouch in addition to the multicentric T-cell lymphoma.

Solute and particle retention in the digestive tract of the Phillip's dikdik (Madoqua saltiana phillipsi), a very small browsing ruminant: biological and methodological implications.

Morphological characteristics of the forestomach, as well as reports of a natural diet that mostly excludes monocots, suggest that dikdiks (Madoqua spp.), among smallest extant ruminants, should have a 'moose-type' forestomach physiology characterised by a low degree of selective particle retention. We tested this assumption in a series of feeding experiments with 12 adult Phillip's dikdiks (Madoqua saltiana phillipsi) on three different intake levels per animal, using cobalt-EDTA as a solute marker and a 'conventional' chromium-mordanted fibre (<2 mm; mean particle size 0.63 mm) marker for the particle phase. Body mass had no influence on retention measurements, whereas food intake level clearly had. Drinking water intake was not related to the retention of the solute marker. In contrast to our expectations, the particle marker was retained distinctively longer than the solute marker. Comparisons with results in larger ruminants and with faecal particle sizes measured in dikdiks suggested that in these small animals, the chosen particle marker was above the critical size threshold, above which particle delay in the forestomach is not only due to selective particle retention (as compared to fluids), but additionally due to the ruminal particle sorting mechanism that retains particles above this threshold longer than particles below this threshold. A second study with a similar marker of a lower mean particle size (0.17 mm, which is below the faecal particle size reported for dikdiks) resulted in particle and fluid retention patterns similar to those documented in other 'moose-type' ruminants. Nevertheless, even this smaller particle marker yielded retention times that were longer than those predicted by allometric equations based on quarter-power scaling, providing further support for observations that small ruminants generally achieve longer retention times and higher digestive efficiencies than expected based on their body size.