RESUMO
BACKGROUND: Antillean manatees (Trichechus manatus manatus) are large herbivorous aquatic mammals living in limited areas of South, Central and North America. As with other aquatic mammals, Antillean manatees can be infected by a variety of protozoan and metazoan parasites, some of them with zoonotic potential, which affect not only their welfare but also population health status. Therefore, we conducted the first epidemiological survey in Colombian free-ranging Antillean manatees to estimate their actual gastrointestinal parasite status. RESULTS: In total, 69 faecal samples were collected from free-ranging individual manatees during ecology field studies in the rivers Carare and San Juan and in two associated wetlands in the Andean region of Colombia. Parasite diversity encompassed six different endoparasite species. The highest prevalence was found for protozoan infections with Eimeria nodulosa (47.8%) and Eimeria manatus-like species (type A, B; 43.4%), followed by Entamoeba sp. (14.49%) and Giardia sp. (1.4%) infections. In addition, infections with the trematode Chiorchis fabaceus were detected at a high prevalence (33.3%). Molecular characterization of sirenian Eimeria species led to the distinction of three species, E. nodulosa and two E. manatus-like species (type A, B). Phylogenetic analyses indicated a host-specific adaptation of sirenian Eimeria species as previously reported for Eimeria species from other mammalian hosts. CONCLUSIONS: This study provides the first record of Antillean manatee infection with Giardia and Entamoeba species in Colombia, representing two important anthropozoonotic parasite genera. This survey should serve as a baseline investigation for future monitoring on parasitic zoonoses in this mammal and encourage for investigations on their impact on both public health and wild manatee welfare.
Assuntos
Animais Selvagens/parasitologia , Enteropatias Parasitárias/veterinária , Doenças Parasitárias em Animais/epidemiologia , Trichechus manatus/parasitologia , Animais , Colômbia/epidemiologia , Eimeria/isolamento & purificação , Entamoeba/isolamento & purificação , Giardia/isolamento & purificação , Enteropatias Parasitárias/epidemiologia , Filogenia , Rios/parasitologia , Trematódeos/classificação , Trematódeos/isolamento & purificação , Zoonoses/epidemiologia , Zoonoses/parasitologiaRESUMO
OBJECTIVE: To determine whether the human spermatozoon is a sufficient stimulus to trigger the release of neutrophil extracellular traps (NETs) in a time- and dose-dependent manner. DESIGN: Experimental study. SETTING: University-based laboratory. PATIENT(S): Semen samples from four men and blood samples from six healthy female donors. INTERVENTION(S): Polymorphonuclear neutrophils (PMN) isolated from peripheral blood were incubated with fresh human spermatozoa for 60, 90, 120, and 180 minutes and at different PMN/sperm concentrations (1:1 [25 × 104], 1:3 [25 × 104:75 × 104], 1:6 [25 × 104:15 × 105], 1:18 [25 × 104:45 × 105]). MAIN OUTCOME MEASURE(S): During coincubation of PMN/sperm, the release of NETs was measured by PicoGreen. Immunofluorescence for histone H3, neutrophil elastase (NE), and myeloperoxidase (MPO) was performed. Different NETs inhibitors were tested: diphenylene iodonium, Suc-Ala- Ala-Pro-Val chloromethyl ketone (CMK), and 4-aminobenzoic acid hydrazide (ABAH) inhibitors of NADPH oxidase, NE, and MPO. Progressive mobility was assessed at increasing doses of neutrophils (1:18 [25 × 104:45 × 105], 6:18 [15 × 105:45 × 105], 9:18 [252 × 104:45 × 105]). RESULT(S): The quantity of NETs increased at the ratio of 1:6 after 2 hours and continued to increase subsequently. A ratio of 1:18 showed significant increases in NETs production at all times. Assessment of the inhibitors showed that CMK and ABAH inhibit NETs formation. Scanning and transmission electron microphotographs and immunofluorescence confirmed NETs formation induced by the spermatozoa. After 1 hour, progressive motility diminished in the two groups with the highest proportion of neutrophils and after 2 hours in all groups exposed to neutrophils. CONCLUSION(S): We show that the stimulus of the human spermatozoon triggers the release of NETs; this response is dose dependent and increases with exposure time. The motility of affected spermatozoa diminishes, suggesting that this interaction on a larger scale would decrease the probability of successful fertilization.