Non-invasive detection of Orthohalarachne attenuata (Banks, 1910) and Orthohalarachne diminuata (Doetschman, 1944) (Acari: Halarachnidae) in free-ranging synanthropic South American sea lions Otaria flavescens (Shaw, 1800).

Respiratory mites of the genera Orthohalarachne and Halarachne (Acari: Halarachnidae) are causative agents of nasopharyngeal/nasopulmonary acariasis in pinnipeds and sea otters. Until now, these endoparasitic mites were mainly diagnosed via necropsies and invasive procedures. So far, non-invasive diagnostic techniques have neither been developed nor applied in free-ranging pinnipeds. In the current study, we aimed to evaluate the halarachnid mite infestation status of free-ranging "urban" South American sea lions Otaria flavescens in the city of Valdivia, Chile. Therefore, non-invasive sampling methods were used in the current study, e. g. by observation-based sampling of freshly expectorated nasal mucus in the animal environment. Further, collection devices were developed for target-oriented sampling of sneezed nasal mucus, including sterile petri dishes and stretched clingfilms mounted on telescopic rods. Applying these techniques, 26 individual sputum samples were collected. 11.5% of sputum samples proved positive for halarachnid larvae (in total, n = 7), which were morphologically identified as Orthohalarachne attenuata (n = 2) or Orthohalarachne diminuata (n = 5). In one of the individual sea lion mucus samples, both Orthohalarachne species were detected, thereby confirming a patent co-infestation in vivo. 16S rDNA-based molecular identification of individual Orthohalarachne spp. larvae confirmed morphological findings. For the first time, we here molecularly characterized Orthohalarachne spp. on the basis of three gene regions [18S, 28S and the internal transcribed spacer 1 (ITS1)]. Overall, current data include the successful application of non-invasive techniques to sample halarachnid mites from free-ranging synanthropic pinnipeds and contribute to the current knowledge on respiratory mites infesting South American sea lions by combining morphological and molecular methods to overcome challenges in species identification. This study should further serve as baseline study and calls for more research on occurrence, biology and health implications of orthohalarachnosis in free-living as well as captive pinnipeds.

Bracken-fern extracts can be clastogenic or aneugenic depending on the tissue cell assay.

The consumption of bracken fern (Pteridium aquilinum) as food is associated with a high incidence of cancer in humans and animals. We investigated the cytogenetic effects of bracken-fern extracts (hexane extract-HE, ethanol extract-EE, hot water extract-HWE and cold water extract-CWE) on chromosomes of peritoneal and bone-marrow cells of Swiss mice. In peritoneal cells, all four treatments (HE, EE, HWE and CWE) induced structural chromosome aberrations, but the EE also induced numerical chromosome aberrations. In bone-marrow cells both HE and CWE induced structural chromosome aberrations; additionally, the number of abnormal metaphases was higher in peritoneum than in bone marrow. We suggest that bracken fern induces cytogenetic damage through DNA strand breaks and affects chromosome segregation.

Differential susceptibility to chromatid breaks induced by bleomycin in sub-fertile and fertile bovines.

The rate of chromatid breaks was studied in cows with a history of sub-fertility by means of a test based on measurement of the average of breaks induced in lymphocytes of peripheral blood cultures. Fourteen female specimens were divided into two groups: fertile and sub-fertile. Peripheral blood lymphocytes were cultured and prepared for cytogenetic analysis. Two types of culture were established for each animal to evaluate the response of peripheral blood lymphocyte cultures to the genotoxic effects of bleomycin. The first culture did not receive bleomycin treatment (spontaneous chromosome aberrations). Our results showed that median breaks per cell (b/c) (+/-semirange) for spontaneous culture of the fertile and sub-fertile animals and bleomycin sensitivity assay for fertile and sub-fertile animals were 0.00+/-0.06, 0.02+/-0.03, 0.08+/-0.05 and 0.22+/-0.09, respectively. There was no significant difference (P>0.05) in the chromosomal breakage in lymphocytes not exposed to bleomycin; however, in comparing the number of chromatid breaks per cell in cultures treated with bleomycin, the sub-fertile group showed a significantly higher (P<0.05) level than the fertile group. These findings have implications both for identifying cattle with less than optimum fertility as well as for providing potential avenues to study the origins of sub-fertility.

Inserçäo de DNA de Trypanosoma cruzi no genoma de célula hospedeira de mamífero por meio de infecçäo / Insertion of Trypanosoma cruzi DNA in the genome of mammal host cell through infection

Observamos a cromatina de formas amstigotad de T. cruzi associada a cromossomos de macrófagos metafásicos obtidos em diversos períodos da infecçäo aguda. O estudo imunocitogenético demonstrou que o material genético inserido naqueles cromossomos era produto do T. cruzi. Pelo teste de hibridaza ao in situ com sonda biotinilada de DNA de T. cruzi, foi confirmada a inserçäo de DNA do protozoário nos cromossomos murinos. A inserçäo seletiva de 3H-DNA do protozoário em alguns cromossomos sugere que podem ocorrer rearranjos transxenogênicos em infecçöes de mamiferos pelo T. cruzi