Combining morphological and molecular data to reconstruct the phylogeny of avian Haemosporida.

The traditional classification of avian Haemosporida is based mainly on morphology and life history traits. Recently, molecular hypotheses have challenged the traditional classification, leading to contradictory opinions on whether morphology is phylogenetically informative. However, the morphology has never been used to reconstruct the relationships within the group. We inferred the phylogeny of avian Haemosporida from 133 morphological characters present in blood stages. We included all species with at least one mitochondrial gene characterized (n = 93). The morphological hypothesis was compared with the one retrieved from mitochondrial DNA (mtDNA) nucleotide sequences and a hypothesis that used a combination of morphological and molecular data (i.e., total evidence). In order to recover the evolutionary history and identify phylogenetically and taxonomically informative characters, they were mapped on the total evidence phylogeny. The morphological hypothesis presented more polytomies than the other two, especially within Haemoproteus. In the molecular hypothesis, the two Haemoproteus subgenera are paraphyletic, and some relationships within Parahaemoproteus were resolved. By combining the morphological and molecular data, we were able to resolve the majority of polytomies and posterior probabilities increased. We identified a unique combination of morphological traits, clearly differentiating avian Haemosporida genera, sub-genera of Leucocytozoon and Haemoproteus, and some Plasmodium sub-genera. Plasmodium had the highest number of synapomorphies. Furthermore, 86% of the species presented a unique combination of taxonomically informative characters. A limiting factor was the mismatch of traits characterized in species descriptions, leading to a morphological matrix with a considerable amount of missing data, particularly for the stages of early young and young gametocytes (67% of all missing data). Characters lacking information for the majority of species included the colour of pigment granules, the cytoplasm appearance, and the presence and dimensions of vacuoles. According to our results, the combination of morphology and mtDNA proved to be a robust alternative to reconstruct the relationships among avian Haemosporida, obtaining a resolution and support similar to that obtained using full mitochondrial genome sequences for over 100 lineages.

Biochemical and hematological evaluations of black howler monkeys (Alouatta pigra) in highly degraded landscapes in Mexico.

BACKGROUND: Loss and fragmentation of Neotropical primates' habitat can alter the health and individual adaptation. Physiological parameters reflect health status and individuals responses to the habitat conditions. METHODS: We captured six wild adult females and six adult males of Alouatta pigra to evaluate their physical condition (body mass, respiratory and heart rate, and rectal temperature), hematology and blood chemistry on fragmentation habitat condition at southeastern Mexico. RESULTS: The males weighed more than females, but the female' temperature, respiratory rate, and heart rate were higher. Low values of hemoglobin, hematocrit, and red blood cells suggest some degree of anemia in these howler monkeys. Low levels of the total protein, albumin, albumin/globulin Ratio, and low levels of blood urea nitrogen suggest that howler monkeys could have protein and carbohydrate deficiencies. CONCLUSIONS: Howler monkeys living under the highly degraded conditions at Balancán, Tabasco, could be showing adaptability by modifying their physiology, hematology, and blood chemistry in response to the conditions of their fragmented landscape.

Contrasting time-based and weight-based estimates of protein and energy intake of black howler monkeys (Alouatta pigra).

Two methods are commonly used to describe the feeding behavior of wild primates, one based on the proportion of time animals spent feeding on specific plant parts ("time-based" estimates) and one based on estimates of the actual amounts of different plant materials ingested ('"weight-based" estimates). However, studies based on feeding time may not be accurate for making quantitative assessments of animals' nutrient and energy intake. We analyzed the diet of two groups of Alouatta pigra living in forest fragments using two different methods (time- and dry weight-based estimates), to explore how these alternative approaches impact estimates of (a) the contribution of each food type to the diet and (b) the macronutrient composition of the diet, including available protein (AP), non-protein energy (NPE), and total energy (TE) intake. We conducted behavioral observations (N = 658 hr and N = 46 full day focal follows), from August 2012 to March 2013. For each feeding bout, we estimated both time spent feeding and actual fresh- and dry-weight consumption by counting the number of food items ingested during the bout. Using time-based estimates, A. pigra showed a predominantly leaf-based diet. In contrast, weight-based estimates described combined a fruit and leaf-based diet. There were no differences between methods when estimating AP intake; however, we found significant differences while estimating NPE and TE intake. Time-based estimates provide us with important information such as the foraging effort spent on food items, trees, or patches, while weight-based estimates may provide more accurate information concerning nutrient and energy intake. We suggest that quantitative estimates of nutrient intake in a primate's diet be based on observations of wet and/or dry weight actually ingested rather than extrapolated from time spent feeding.

Organochlorine Pesticides in the Ferruginous Pygmy Owl (Glaucidium brasilianum) in Chiapas, Mexico.

Concentrations of organochlorine pesticides were quantified in samples of feathers (n = 17) and blood (n = 15) of the ferruginous pygmy owl (Glaucidium brasilianum). The individuals were captured near the Protected Natural Area Cerro Sonsonate, Chiapas, Mexico, between February and June 2014. In both tissues, pesticides belonging to seven organochlorine chemical families were detected. However, the organochlorine pesticide concentrations differed between feathers and blood. The highest concentrations of hexachlorocyclohexanes were found in feathers (0.63 ± 0.89 µg/g), whereas the highest concentrations of ΣDrines were found in blood (0.31 ± 0.47 µg/mL). By using the summed concentrations for each of the seven families of pesticides found in feathers, we did not find any significant correlation between the pesticides and pectoral muscle or body weight (p > 0.15). The ΣDDT group was the only pesticide family that showed a positive correlation with owl body weight (r = 0.60, p = 0.05); the concentrations of these pesticides were also high in feather and blood tissues (r = 0.87, p = 0.02). Our results confirm that ferruginous pygmy owls in the study area are exposed to these pesticides.

Cascading impacts of anthropogenically driven habitat loss: deforestation, flooding, and possible lead poisoning in howler monkeys (Alouatta pigra).

To construct informed conservation plans, researchers must go beyond understanding readily apparent threats such as habitat loss and bush-meat hunting. They must predict subtle and cascading effects of anthropogenic environmental modifications. This study considered a potential cascading effect of deforestation on the howler monkeys (Alouatta pigra) of Balancán, Mexico. Deforestation intensifies flooding. Thus, we predicted that increased flooding of the Usumacinta River, which creates large bodies of water that slowly evaporate, would produce increased lead content in the soils and plants, resulting in lead exposure in the howler monkeys. The average lead levels were 18.18 ± 6.76 ppm in the soils and 5.85 ± 4.37 ppm in the plants. However, the average lead content of the hair of 13 captured howler monkeys was 24.12 ± 5.84 ppm. The lead levels in the animals were correlated with 2 of 15 blood traits (lactate dehydrogenase and total bilirubin) previously documented to be associated with exposure to lead. Our research illustrates the urgent need to set reference values indicating when adverse impacts of high environmental lead levels occur, whether anthropogenic or natural, and the need to evaluate possible cascading effects of deforestation on primates.

Noninvasive collection of fresh hairs from free-ranging howler monkeys for DNA extraction.

The use of noninvasive collected samples as source of DNA in studies of wild primate populations has increased in recent years. Fresh-plucked hairs represent an important source of DNA, with relatively high quality and concentration. In this study, we describe a low-cost noninvasive technique for collecting fresh-plucked hairs used to obtain DNA samples from free-ranging black howler monkey populations (Alouatta pigra). We designed and manufactured darts made of wooden dowels, with the anterior part smeared with glue, which were projected with blowpipes to trap howler monkey hairs. All of the materials to make the darts are inexpensive and are available locally. We collected 89 samples from 76 individuals residing in 15 troops, and the total number of hairs obtained was 754. We found no differences in the number of hairs collected among sex-age classes or among localities but the percentage of darts recovered with sample varied among localities. Preliminary results indicate that over 96% of samples yielded DNA suitable for polymerase chain reaction-based microsatellite marker analysis. The technique proved successful for collecting fresh-plucked hairs of free-ranging black howler monkeys without any trauma to the animals and can be easily adapted to obtain samples from other wild primate and mammal species.