Prevalence and molecular characterization of Anaplasmataceae agents in free-ranging Brazilian marsh deer (Blastocerus dichotomus)

Anaplasmataceae organisms comprise a group of obligate intracellular gram-negative, tick-borne bacteria that can infect both animals and humans. In the present work we investigate the presence of Ehrlichia, Anaplasrna, and Neorickettsia species in blood samples from Brazilian marsh deer (Blastocerus dichotomus), using both molecular and serologic techniques. Blood was collected from 143 deer captured along floodplains of the Parana River, near the Porto Primavera hydroelectric power plant. Before and after flooding, marsh deer were captured for a wide range research program under the financial support of São Paulo State Energy Company (CESP), between 1998 and 2001. Samples were divided into four groups according to time and location of capture and named MS01 (n = 99), MS02 (n = 18) (Mato Grosso do Sul, before and after flooding, respectively), PX (n = 9; Peixe River, after flooding), and AGUA (n = 17; Aguapei River, after flooding). The seroprevalences for Ehrlichia chaffeensis and Anaplasma phagocytophilum were 76.76% and 20.2% in MS01, 88.88% and 5.55% in MS02, 88.88% and 22.22% in PX, and 94.12% and 5.88% in AGUA, respectively. Sixty-one animals (42.65% of the total population) were PCR-positive for E. chaffeensis PCR (100.0% identity based on 16S rRNA, dsb, and groESL genes). Seventy deer (48.95% of the total population) were PCR-positive for Anaplasma spp. (99.0% of identity with A. platys, and in the same clade as A. phagocytophilum, A. bovis, and A. platys based on 16S rRNA phylogenetic analysis). Our results demonstrate that Brazilian marsh deer are exposed to E. chaffeensis and Anaplasma spp. and may act as reservoirs for these rickettsial agents, playing a role in disease transmission to humans and other animals. (c) 2012 Elsevier Ltd. All rights reserved.

Cytogenetic analysis of the Marsh Deer, Blastocerus dichotomus (Mammalia, Cervidae)

The chromosomic constitution of the Marsh Deer (Blastocerus dichotomus) was studied in 18 males and 18 females, mainly from the Tiete river basin in Sao Paulo State, Brazil. The species diploid number was determined to be 66 chromosomes and the fundamental number (FN), 74. The X and the Y were the largest and the smallest chromosome, respectively. Large amounts of the constitutive heterochromatin marked by the C band were located in the centromeric region of all the acrocentric chromosomes. The first chromosome pair was not marked and the second and third pairs showed weak centromeric markings. The X chromosome showed two strong telomeric markings while the Y was C band negative. Chromosomes four and five were the NOR carriers. Polymorphism for this band was observed in pair four. The results of this study are in agreement with other reports in the literature, in spite of the different origin of the animals.

Ticks (Acari: Ixodidae) on Wild Marsh-Deer (Blastocerus dichotomus) from Southeast Brazil: Infestations before and after Habitat Loss

The lake from Porto-Primavera hydroelectric power station inundated an area of 2,200 km2 at the border of São Paulo and Mato-Grosso do Sul States, Brazil. Infestations by ticks were evaluated on 135 marsh deer, Blastocerus dichotomus (Illiger), captured before and after inundation. Ticks were collected for identification, and infestation level of animals was assessed by scoring. Deer were divided into four groups according to capture location and temporal relation to the inundation. Groups 1, 2, and 3 were captured before inundation. Group 4 was captured after inundation. Four tick species were found: Amblyomma cajennense (F.), Amblyomma triste Koch, Anocentor nitens (Neumann), and Boophilus microplus (Canestrini). Groups 1, 2, 3, and 4 had 30, 45, 100, and 96%, respectively, of animals carrying B. microplus ticks. A. triste was observed on 16, 22, 22, and 88% of animals from groups 1,2,3, and 4, respectively. A. nitens and A. cajennense were observed only on group 4, on 32 and 16% of the animals, respectively. Groups 1 and 2 had only 4.8 and 6.1% of animals with high infestation levels, respectively, and no ticks on 46.8% and 45.5% of the animals, respectively. Conversely, groups 3 and 4 lacked noninfested animals and had high infestation levels on 77.8 and 50% of deer, respectively. Marsh area shrinkage was blamed for higher infestation levels on deer from groups 3 and 4. The widespread presence of A. triste on marsh deer, a Neotropical tick species, raises the possibility of a natural host-parasite relationship.

Tuberculosis survey of free-ranging marsh deer (Blastocerus dichotomus) in Brazil

Esophageal-pharyngeal fluids from 53 free-ranging marsh deer (Blastocerus dichotomus) captured for a research program in the state of Mato Grosso do Sul, Brazil, were assayed for tuberculosis. Total DNA was extracted, amplified by polymerase chain reaction using specific primers for Mycobacterium tuberculosis complex (M. tuberculosis, M. bovis, M. microti, and M. africanum), and observed by agarose gel electrophoresis stained with ethidium bromide. All samples were negative. This, along with necropsy and histopathology data, suggests that these animals are not shedding and probably do not have active disease.

Natural infection by Paramphistomoidea Stiles and Goldberger, 1910 trematodes in wild Marsh Deer (Blastocerus dichotomus Illiger, 1815) from Sérgio Mottas's hydroelectric power station flooding area.

Studies on helminthfauna of marsh deer Blastocerus dichotomus Illiger, 1815 are rare, although helminthic diseases are an important cause of mortality in these animals. Fifteen male and female adult marsh deer from Sergio Motta's hydroelectric power station flooding area at Paraná River which died during the capture and quarantine procedures, between 1998 and 1999, were necropsied. Three trematodes species, Paramphistomum cervi, Balanorchis anastrofus and Zygocotyle lunatum, all belonging to superfamily Paramphistomoidea, were identified. The obtained trematodes were identified, counted and their respectives descriptors of infection were determined. All necropsied animals were infected by helminths. Paramphistomum cervi was the most prevalent species, while Zygocotyle lunatum was found in only one animal.

Transferability of microsatellite loci from Cervidae species to the endangered Brazilian marsh deer, Blastocerus dichotomus

Blastocerus dichotomus, the marsh deer, is the largest Brazilian Cervidae species. The species is endangered because of hunting and loss of its natural habitat, i.e., flood plain areas, because of hydroelectric power station construction and agricultural land expansion. In the present study, we tested 38 microsatellite loci from four Cervidae species: Odocoileus virginianus (7), Rangifer tarandus (17), Capreolus capreolus (7), and Mazama bororo (7). Eleven loci showed clear amplification, opening a new perspective for the generation of fundamental population genetic data for devising conservation strategies for B. dichotomus. © FUNPEC-RP.

Genética da reintrodução: o caso do cervo-do-pantanal (Blastocerus dichotomus) na Estação Ecológica de Jataí

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Aglutininas contra Leptospira spp em cervos-do-pantanal (Blastocerus dichotomus) na área afetada pela construção da usina hidrelétrica Sérgio Motta

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identificação e Prevalência de Agentes Rickettsiais (Rickettsiales: Anaplasmataceae) em Cervo-do-pantanal (Blastocerus dichotomus) utilizando métodos sorológico e molecular

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

Determinação dos perfis de estrógenos e progestinas fecais durante o ciclo estral, gestação e período pós-parto em cervos-do-pantanal (Blastocerus dichotomus) em cativeiro

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

Severe population decline of marsh deer, Blastocerus dichotomus (Cetartiodactyla: Cervidae), a threatened species, caused by flooding related to a hydroelectric power plant

Balancing power production and environmental conservation can be problematic. The objective of this study was to investigate the abundance of marsh deer in the Paraná River Basin, above the Sergio Motta (Porto Primavera) Dam, before and after the impact of the dam closure. A fixed-wing, flat window aircraft was used to survey study transects. Observations were recorded based on the distance sampling line transect method, assuming that the detection probability decreases with increased distance. The abundance of marsh deer in the survey region prior to flooding was estimated to be 974 individuals (CV = 0.23). The overall abundance dropped from 974 to 444 (CV = 0.26) individuals after flooding, an overall reduction of 54%. This reduction can be attributed to the direct impact of the flooding process, but it was likely exacerbated by indirect effects, such as increased disease, hunting, and reduction in food availability. Prior to flooding, the marsh deer was distributed widely throughout the dam's catchment area; however, the marsh deer habitat was almost completely destroyed by the flooding process. This situation highlights the need to implement management strategies that ensure the survival of the remaining fragmented population.

Morfologia do estômago do cervo-do-pantanal (Blastocerus dichotomus, Illiger 1815)

Based on the lack of information regarding the morphology of marsh deer, this work aims to describe some morphological aspects of the gastric chamber in this species, collaborating with future investigations, mainly related to rational handling in this cervid. This work aimed to describe the morphology of the gastric chamber of the marsh deer, characterizing the external and internal macroscopical details and the microscopical architecture of these structures by light microscopy. Macroscopically, the marsh deer stomach is formed by the rumen, reticulum, omasum and abomasum similar to the domestic ruminants. Microscopically, rumen and abomasum are similar to the domestic ruminants. The reticulum and the omasum, however, present specific characteristics such as keratin on the top of the reticulum, small epithelial projections and omasum folds covered with discrete papillae.

Infecções naturais em cervídeos (Mammalia: Cervidae) procedentes dos Estados do Mato Grosso do Sul e São Paulo, por nematódeos Trichostrongyloidea Cram, 1927

No período compreendido entre 1985 e 1996 foram necropsiados, para pesquisa de helmintos, 42 cervídeos, sendo sete Mazama americana, 16 M. gouazoubira, 13 Ozotoceros bezoarticus e seis Blastocerus dichotomus. Desses animais, foram colhidos 14.426 nematódeos Trichostrongyloidea, sendo 13.281 (92,06%) parasitos de abomaso e 1.145 (7,94%), de intestino delgado. Nesses órgãos, foram identificadas seis espécies de nematódeos: Haemonchus contortus, H. similis, Trichostrongylus axei, T. colubriformis, Cooperia punctata e C. pectinata. Todos os animais apresentaram infecções helmínticas por uma ou mais espécies, ocorrendo grande variação na intensidade de infecção (1 a 4.345 nematódeos). Ainda com relação à intensidade de infecção, os dados expressavam valores menores que 100 parasitos em 25 (59,52%) animais. Os valores mais altos de intensidade média das infecções foram observados em M. gouazoubira (596,37 helmintos) e em O. bezoarticus (331), e os menores, em M. americana (17,57) e B. dichotomus (75,5). Os dados mais expressivos de intensidade de infecção, abundância e prevalência foram observados para Haemonchus (larvas de 4º estágio), H. contortus, H. similis e T. axei. O gênero Haemonchus foi constatado em 35 animais, com prevalência de 83,33%; apresentou carga parasitária de 11.616 exemplares, representando 80,52% dos nematódeos verificados, sendo a maioria (8.903) constituída por formas imaturas. Por outro lado, H. similis foi a espécie predominante nas infecções e, portanto, a que apresentou maiores valores de abundância. Verificou-se o gênero Trichostrongylus em 24 (57,14%) animais, com carga parasitária de 2.444 exemplares, sendo 1.665 espécimes de T. axei, que representou 11,54% da carga parasitária obtida. As seis espécies de vermes identificadas nos cervídeos são comuns aos ruminantes domésticos nos Estados de São Paulo e Mato Grosso do Sul e dessa maneira não se observou nenhuma espécie de Trichostrongyloidea exclusiva dos cervídeos.

Gastro-intestinal transit time in South American deer

The study of diet and physiological peculiarities of the digestive system of neotropical deer is not well known and the literature shows inconsistencies. To better understand the digestive system of these mammals the difference in the gastro-intestinal transit time of four species of neotropical deer (Mazama americana, Mazama gouazoubira, Mazama nana, Blastocerus dichotomus), kept ill captivity, was evaluated. Four plants (Neonotonia wightii, Morus albans, Medicago sativa, Leucaena leucocephala) were utilized and two variables were measured, mean time of the beginning of the elimination (BE) and mean time of permanence (TP). The results obtained for BE indicated similarity among the deer species, with significant differences between M. gouazoubira (mean = 13.62 hr) and M. Americana (mean = 19.25 hr). For the plants, the BE was faster with N. wightii and L. leucocephala, and slower for M. sativa. The TP results for B. dichotomus showed longer time when compared to the other species, whereas M. gouazoubira had a lower permanence. Overall, N. wightii had the highest retention time in the digestive tract of all the deer species studied. Associated with this observation, N. wightii had the highest quantity of plant fiber of the plants tested. In a similar fashion M. sativa showed the lowest TP in the digestive tract of the deer and had the lowest quantity of acid detergent fiber. The data from this study showed that, within species, the shape of the excretion curve of the plants was similar when the animals consumed N. wightii or M. sativa. Blastocerus dichotomus and M. gouazoubira had the highest and the lowest gastro-intestinal transit time, respectively. This suggests that these species characterize different abilities to digest high fiber food, and consequently, represent the two extremities in the morphophysiological adaptation within the deer species evaluated. This information is vital because it is important to know the digestive physiology to define the diet of captive animals, particularly regarding the quantity and quality of fiber. Zoo Biol 25:47-57, 2006. (C) 2005 Wiley-Liss, Inc.

Prevalence and Molecular Characterization of Mycoplasma ovis in Selected Free-Ranging Brazilian Deer Populations

Mycoplasma ovis is a hemoplasma that may cause anemia and mortality in small ruminants. Our aim was to determine whether M. ovis infects populations of free-ranging deer in Brazil. Bully coat samples from 64 Blastocerus dichotomus from Porto Primavera, 18 Ozotocerus bezoarticus from Pantanal, and 21 O. bezoarticus from Emas National Park were tested. Using a M. ovis PCR protocol to amplify extracted DNA, 46/64 (72%) of deer froth Porto Primavera, 10/18 (56%) from Pantanal, and 4/21 (19%) from Emas National Park were positive, giving an overall positive rate of 58% for hemoplasma in these wild deer. Sequencing and phylogenetic analysis of the 168 rRNA gene revealed 3 genetically distinct hemoplasmas including M. ovis, 'Candidatus Mycoplasma erythrocervae', and a hemoplasma most closely related to M. ovis. Phylogenetic analysis of the 23S rRNA gene from selected sequences confirmed these relationships.