Amplifiability of mitochondrial, microsatellite and amelogenin DNA loci from fecal samples of red brocket deer Mazama americana (Cetartiodactyla, Cervidae)

We tried to amplify mitochondrial, microsatellite and amelogenin loci in DNA from fecal samples of a wild Mazama americana population. Fifty-two deer fecal samples were collected from a 600-ha seasonal semideciduous forest fragment in a subtropical region of Brazil (21°20′, 47°17′W), with the help of a detection dog; then, stored in ethanol and georeferenced. Among these samples 16 were classified as fresh and 36 as non-fresh. DNA was extracted using the QIAamp® DNA Stool Mini Kit. Mitochondrial loci were amplified in 49 of the 52 samples. Five microsatellite loci were amplified by PCR; success in amplification varied according to locus size and sample age. Successful amplifications were achieved in 10/16 of the fresh and in 13/36 of the non-fresh samples; a negative correlation (R = -0.82) was found between successful amplification and locus size. Amplification of the amelogenin locus was successful in 22 of the 52 samples. The difficulty of amplifying nuclear loci in DNA samples extractedfrom feces collected in the field was evident. Some methodological improvements, including collecting fresh samples, selecting primers for shorter loci and quantifying the extracted DNA by real-time PCR, are suggested to increase amplification success in future studies. © FUNPEC-RP.

Fecal progestins during pregnancy and postpartum periods of captive red brocket deer (Mazama americana)

This study aimed to validate the enzyme immunoassay (EIA) for fecal progestin quantification of the species Mazama americana, define its excretion profile during periods of gestation and postpartum and determine the gestation period and resumption of postpartum ovarian activity in this species in captivity Fecal samples were collected twice a week during gestation and every day in the postpartum period, and analyzed using EIA The mean concentrations (±SEM) of fecal progestins during gestation were 2180.0 ± 299.1 ng/g in early pregnancy (week 1-11), 3271.4 ± 406.9 ng/g in middle pregnancy (week 12-22) and 5592.0 ± 1125.8 ng/g in late pregnancy (week 23-32) The gestation period determined for the species was 220.9 ± 1.2 days The concentration of progestins reached its peak prior to parturition and returned to baseline levels in 4 ± 0.31 days after parturition In the postpartum period, the mean concentrations of fecal progestins were 1564.2 ± 182.6 ng/g in the interval between parturition and resumption of ovarian activity, 469.8 ± 24.5 ng/g in the inter-luteal phase and 2401.7 ± 318.5 ng/g during the luteal phase, such that the postpartum period and the luteal phase differed from the inter-luteal phase Fecal progestin profiling permitted the detection of ovulation 26.9 ± 3.4 days after parturition in all the hinds studied and estimation of the mean duration of the estrous cycle, 21.3 ± 1.1 days Analysis established that concentrations of progestins above 3038.76 ng/g diagnosed pregnancy, a value determined from the week 12 of gestation Moreover, the quantification of fecal progestins by EIA proved to be an important tool for noninvasive endocrine monitoring and to obtain reproductive data on the species M americana in captivity © 2013 Elsevier B.V.

Meiotic pairing of B chromosomes, multiple sexual system, and Robertsonian fusion in the red brocket deer Mazama americana (Mammalia, Cervidae)

Deer species of the genus Mazama show significant inter and intraspecific chromosomal variation due to the occurrence of rearrangements and B chromosomes. Given that carriers of aneuploidies and structural rearrangements often show anomalous chromosome pairings, we here performed a synaptonemal complex analysis to study chromosome pairing behavior in a red brocket deer (Mazama americana) individual that is heterozygous for a Robertsonian translocation, is a B chromosome carrier, and has a multiple sex chromosome system (XY1Y2). The synaptonemal complex in spermatocytes showed normal chromosome pairings for all chromosomes, including the autosomal and sex trivalents. The electromicrographs showed homology among B chromosomes since they formed bivalents, but they also appeared as univalents, indicating their anomalous behavior and non-Mendelian segregation. Thus, synaptonemal complex analysis is a useful tool to evaluate the role of B chromosomes and rearrangements during meiosis on the intraspecific chromosomal variation that is observed in the majority of Mazama species. © FUNPEC-RP.

Effect of sex and seasons of the year on hematologic and serum biochemical variables of captive brown brocket deer (Mazama gouazoubira)

The Brown brocket deer (Mazama gouazoubira) is the most common free-living and captive deer in South America, especially in Brazil, and has great ecological and scientific significance. However, data on hematological and biochemical parameters in brown brocket deer are scarce. The goal of this study was to establish reference ranges for hematological and biochemical parameters of Mazama gouazoubira, comparing differences during the seasons of the year and between sex. Blood samples from ten adult healthy brown brocket deer (6 female and 4 male) were collected during daytime, monthly, during 12 months. The animals were maintained in individual stable, protected from noise and fed ad libitum with commercial ration and green fodder. For blood collection, animals were submitted to physical restrain for no longer than 2 minutes. The following parameters were determined: red blood cell count (RBC), haemoglobin concentration, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), white blood cell count (WBC), platelet count, enzyme activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) and serum levels of alkaline phosphatase (ALP), creatine kinase (CK), total protein (TP), albumin, cholesterol, total calcium, ionic calcium, sodium, potassium, magnesium, triglycerides, creatinine and urea. Values were compared according to season and sex. RBC count, WBC count and MCV suggested seasonal influence. Haemoglobin concentration, PCV and MCV were influenced by sex. Serum concentration of total calcium, ionic calcium, sodium, potassium and magnesium were influenced by season. Serum magnesium was also influenced by sex. The blood parameters herein reported may be useful as reference values for diagnostic and prognostic purposes in captive brown-brocket deer.

Comparing two different superovulation protocols on ovarian activity and fecal glucocorticoid levels in the brown brocket deer (Mazama gouazoubira)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The role of chromosome variation in the speciation of the red brocket deer complex: the study of reproductive isolation in females

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

PHYSIOLOGIC EFFECTS OF THREE DIFFERENT PROTOCOLS OF ISOFLURANE ANESTHESIA IN CAPTIVE BROWN BROCKET DEER (MAZAMA GOUAZOUBIRA)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of laparoscopy on the cardiorespiratory system of brown brocket deer (Mazama gouazoubira) anesthetized with ketamine/ xylazine/ midazolam combination and isoflurane

A laparoscopia ainda é pouco utilizada como ferramenta para técnicas de reprodução assistida em cervídeos sul-americanos, não havendo informações sobre seus efeitos e protocolos anestésicos seguros para sua realização. Objetivaramse avaliar as possíveis alterações na freqüência cardíaca (FC), respiratória (FR), saturação de oxihemoglobina (SpO2) e temperatura retal (TR) durante a laparoscopia para visualização dos órgãos reprodutivos de seis fêmeas de veado-catingueiro (Mazama gouazoubira) anestesiadas com a associação cetamina (5mg/kg), xilazina (0,3mg/kg), midazolam (0,5mg/kg) e isofluorano. Cada animal, após anestesiado, foi posicionado em decúbito dorsal para realização de duas laparoscopias com insuflação abdominal de CO2 (14,2 ± 2,39mmHg; M ± EPM) com intervalo de 40 dias. Para avaliar os principais eventos da laparoscopia, esta foi dividida em três períodos: animal sem insuflação abdominal (P1), com insuflação abdominal (P2) e insuflação abdominal com os quadris elevados a 45º (P3). O controle foi realizado após 40 dias da última laparoscopia, para isto, cada animal foi novamente anestesiado e mantido em decúbito dorsal por um período de tempo igual ao tempo médio de duração das anestesias realizadas nas laparoscopias, sem que o procedimento laparoscópico fosse realizado. O tempo de anestesia dos controles foi também dividido em P1, P2 e P3, respeitando o tempo médio de duração de cada um destes períodos das laparoscopias. Para análise dos dados foi usado o teste de análise de variância (ANOVA) seguido do teste de Tukey e valores de P<0,05 considerados significativos. Não houve diferença significativa nos parâmetros estudados em nenhum dos períodos estabelecidos para o controle e laparoscopia. Porém, a FR média entre P1 (38,8 ± 4,42) e P3 (32,7 ± 4,81) e a TR média entre P1 (38,2ºC ± 0,17), P2 (37,6ºC ± 0,19) e P3 (37,0ºC ± 0,21) variaram significativamente, independente da laparoscopia. Tais dados permitiram concluir que a laparoscopia não promoveu alterações significativas nos parâmetros avaliados, embora o protocolo anestésico utilizado tenha contribuído para redução da temperatura retal resultando em risco de hipotermia durante a anestesia.

Influence of husbandry systems on physiological stress reactions of captive brown brocket (Mazama gouazoubira) and marsh deer (Blastocerus dichotomus)-noninvasive analysis of fecal cortisol metabolites

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Different responses to doxorubicin-induced chromosome aberrations in Brazilian deer species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Proteinograma sérico de veados-catingueiro (Mazama gouazoubira) criados em cativeiro obtido por eletroforese em gel de agarose e de poliacrilamida (SDS PAGE)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Non invasive methods for genetic analysis applied to ecological and behavioral studies in Latino-America

Documenting the presence and abundance of the neotropical mammals is the first step for understanding their population ecology, behavior and genetic dynamics in designing conservation plans. The combination of field research with molecular genetics techniques are new tools that provide valuable biological information avoiding the disturbance in the ecosystems, trying to minimize the human impact in the process to gather biological information. The objective of this paper is to review the available non invasive sampling techniques that have been used in Neotropical mammal studies to apply to determine the presence and abundance, population structure, sex ratio, taxonomic diagnostic using mitochondrial markers, and assessing genetic variability using nuclear markers. There are a wide range of non invasive sampling techniques used to determine the species identification that inhabit an area such as searching for tracks, feces, and carcasses. Other useful equipment is the camera traps that can generate an image bank that can be valuable to assess species presence and abundance by morphology. With recent advances in molecular biology, it is now possible to use the trace amounts of DNA in feces and amplify it to analyze the species diversity in an area, and the genetic variability at intraspecific level. This is particularly helpful in cases of sympatric and cryptic species in which morphology failed to diagnose the taxonomic status of several species of brocket deer of the genus Mazama.

Density and population size of mammals introduced on a land-bridge island in southeastern Brazil

The introduction of alien species is one of the main threats to the conservation of native species, especially in island ecosystems. Here, we report on the population growth of 15 species of mammals introduced in 1983 on the island of Anchieta, an 828 ha land-bridge island in southeastern Brazil. We estimated the density of mammals through 296 km of line transect census. Five species introduced became extinct (coypu, brocket deer, six-banded armadillo, nine-banded armadillo, maned three-toed sloth); six became over-abundant (marmoset, coati, agouti, seven-banded armadillo, and capybara); one has a stable population (capuchin monkey). Anchieta Island has the highest density of mammals in the entire Atlantic forest (486.77 ind/km(2)), especially nest predators (232.83 ind/km(2)) and herbivores (253.58 ind/km(2)). Agoutis (Dasyprocta spp.) and marmosets (Callithrix penicillata) were, by far, the species with the highest population growth. The high density of mammals in this island may have strong consequences for plant recruitment and bird diversity.

Evaluation of semen characteristics of the species Mazama americana in captivity

In recent years the concept of genomic resource banks has grown as a way of maintaining the genetic variability of populations, while the quality of cryopreservation of the gametes determines the effectiveness of such banks. However, the absence of basic knowledge regarding the physiology of species and their semen characteristics hampers the establishment of reproductive biotechnologies. Thus, this paper aimed to determine certain physicochemical (volume, colour, appearance, pH and osmolarity) and microscopic characteristics (mass movement, motility, vigour, concentration, and sperm morphology and morphometry) of semen of the species Mazama americana. To achieve this, five males of the species were used, and three semen samples per buck (electroejaculation) were collected at intervals of 2 weeks. The volume, pH and osmolarity of the ejaculate were 0.39 ± 0.14 mL, 6.90 ± 0.74 and 297.74 ± 19.10 mOsm/kg, respectively, while the values obtained for mass movement, motility, vigour and concentration were 3.33 ± 0.82; 69.6 ± 8.92%; 3.53 ± 0.50, and 244.07 ± 98.65 × 107/mL, respectively. Regarding the colour of the ejaculate, five samples were classified as ivory, two as yellowish, two as whitish and six as white. Regarding appearance, seven samples were considered creamy and eight, milky. Morphology was analysed in a humid chamber under phase contrast microscopy and 73.50 ± 5.57% of cells presented normal morphology, 8.37 ± 3.15% presented major defects and 18.13 ± 6.46% presented minor defects. To determine sperm morphometry, an optical microscope (Leica DM 5000B) and the Leica Qwin image analyser program were used, resulting in 8.09 ± 0.40, 4.65 ± 0.30, 2.81 ± 0.44 and 30.25 ± 3.02 m for length, largest width, smallest width and area, respectively. Copyright © CSIRO 2013.