This study was supported by the World Health Organization (H9/TSA/037), the National Research Council of Argentina (PIP 2009-290), the National Agency for Scientific and Technological Promotion of Argentina (PICT 2011, 2023) and the Rene Baron Foundation to P.S.C. and by the MEXT of Japan to M.I. The authors declare that there are no conflicts of interest.

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Llama production in Argentina has increased, as the international interest in breeding this type of animals has grown in the last years. Considering the great polymorphism that llama spermatozoa present at evaluation using light microscopy, the aim of this study was to objectively evaluate llama sperm head morphometry using digital morphometric analysis. Five ejaculates from each of eight males were obtained to evaluate morphometric parameters of 8000 sperm heads stained with Tinción 15(®). The following average results were obtained for each parameter: size parameters: area 20.09 μm(2), length 6.60 μm, width 4.14 μm, equivalent circle diameter 5.06 μm, curve length 5.79 μm and curve width 3.48 μm; boundary parameters: perimeter 18.54 μm and convex perimeter 17.34 μm; and shape parameters: roundness 1.28 and elongation 1.59. Morphometric parameters of sperm head were compared between ejaculates of the same male and between males. Significant differences between ejaculates of the same male were found for all parameters evaluated (P < 0.01). Significant differences between males were found for all morphometric parameters (P < 0.01) except for curve length, curve width and perimeter. The differences detected would indicate that there is not a single morphometric pattern for Lama glama sperm head, because parameter values cannot be standardised.

Epididymal sperm protein CRISP1 has the ability to both regulate murine CatSper, a key sperm calcium channel, and interact with egg-binding sites during fertilization. In spite of its relevance for sperm function, Crisp1-/-mice are fertile. Considering that phenotypes can be influenced by the genetic background, in the present work mice from the original mixed Crisp1-/- colony (129/SvEv*C57BL/6) were backcrossed onto the C57BL/6 strain for subsequent analysis of their reproductive phenotype. Whereas fertility and fertilization rates of C57BL/6 Crisp1-/- males did not differ from those reported for mice from the mixed background, several sperm functional parameters were clearly affected by the genetic background. Crisp1-/- sperm from the homogeneous background exhibited defects in both the progesterone-induced acrosome reaction and motility not observed in the mixed background, and normal rather than reduced protein tyrosine phosphorylation. Additional studies revealed a significant decrease in sperm hyperactivation as well as in cAMP and protein kinase A (PKA) substrate phosphorylation levels in sperm from both colonies. The finding that exposure of mutant sperm to a cAMP analog and phosphodiesterase inhibitor overcame the sperm functional defects observed in each colony indicated that a common cAMP-PKA signaling defect led to different phenotypes depending on the genetic background. Altogether, our observations indicate that the phenotype of CRISP1 null males is modulated by the genetic context and reveal new roles for the protein in both the functional events and signaling pathways associated to capacitation.

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We evaluated a number of lipophilic dyes and fluorochromes, including oxazone and thiazone derivatives of oxazine and thiazine dyes, scintillator agents, a carotenoid and a metal-porphyrin complex for visualization of lipid droplets within aldehyde fixed cultured HeLa and BGC-1 cells. Observation under ultraviolet, blue or green exciting light revealed selective fluorescence of lipid droplets, particularly after treatment with aqueous solutions of Nile blue and brilliant cresyl blue oxazones, toluidine blue thiazone, or propylene glycol solutions of canthaxanthin, ethyl-BAO, and ZnTPyP. Mounting in water was required to maintain the fluorescence of lipids; the use of glycerol, Mowiol or Vectashield was not adequate. The effect of dye structure on staining intensity was assessed with the aid of numerical structure parameters modeling lipophilicity (HI and log P), overall size (MW) and the size of the conjugated system (conjugated bond number; CBN). The best stains for lipid droplets were relatively lipophilic (HI > 4.0, log P > 5.0), of small size overall (MW < 370), with small conjugated systems (CBN < 24), and not significantly amphiphilic. The two hydrophobic-hydrophilic parameters (the classic log P and the hydrophobic index, HI; values calculated by molecular modeling software) were highly correlated; however, HI was a more suitable hydrophobicity index for the dyes studied here.

Heterochronic events in the ontogeny of Columba livia, Coturnix coturnix, and Gallus gallus domesticus ¤ Eventos heterocrónicos en la ontogenia de Columba livia, Coturnix coturnix y Gallus gallus domesticus Eventos heterocrônicos na ontogenia da Columba livia, Coturnix coturnix e Gallus gallus domesticus SummaryBackground: a series of events take place in a precise spatial and temporal context during the development of any organism. Typically, certain ontogenetic processes are consistent with the proper completion of previous events. Objective: to identify possible heterochronic events that may relate to altricial and precocious development patterns. Methods: we analyzed the ontogeny of three species of birds with two different development models: Columba livia (semialtricial 2), Coturnix coturnix, and Gallus gallus domesticus (precocial 2). The starts and endings of thirteen morphological events were compared, from hour 16 of incubation to the time of hatching. Results: while no differences in the sequence of developmental events were found, the events of the maturation stage in the altricial kind started earlier compared to the precocial species. Ontogenetic acceleration events in C. livia and C. coturnix explain how these species reach a level of morphological development similar to that of G. gallus domesticus, but with shorter incubation period. Conclusion: the results provide information not considered in the literature of the specialty about heterochronic events in early developmental stages of poultry.

Poxvirus skin disease has been reported in several species of cetaceans, principally in odontocetes, and a single report in mysticetes. Southern right whales Eubalaena australis in Peninsula Valdes, Argentina, show a variety of skin lesions of unknown etiology, and the number of these lesions has increased in recent years. Samples from dead whales were taken in order to establish the etiology of these lesions. One calf and one adult presented ring-type lesions, characterized by a circumscribed and slightly raised area of skin. Lesions were histologically characterized by the presence of microvesicles and vacuolated cells in the stratum spinosum, along with hyperplasia of the stratum corneum and eosinophilic inclusion bodies in the cytoplasm of the epithelial cells. Transmission electron microscopy showed aggregations of virions with typical poxvirus morphology. PCR of cetacean poxvirus (CPV) DNA polymerase, DNA topoisomerase I and parapoxvirus DNA polymerase gene fragments was done, and confirmed the presence of poxvirus in one sample. Phylogenetic analysis showed that the detected poxvirus belongs to the CPV-2 group. This is the first confirmed report of poxvirus in southern right whales in Argentina.