Brucella Infection Investigation in Cetaceans and Manatees in Northeast Brazil.

Among the bacterial infections that impair the health status of marine mammals, those caused by Brucella spp. are the most reported worldwide. Brucella infections in marine mammals can result in acute or chronic disease and are associated with variable clinical outcomes, depending on the organ involved during the infectious process, infection route, host immunity, and strain pathogenicity. Asymptomatic infections may also occur. The current study expands the investigation of Brucella infection in northeast Brazil by analyzing 19 dead, stranded cetaceans and 52 Antillean manatees Trichechus manatus manatus. The manatees included 8 dead, captive manatees and 44 live specimens, of which 10 were analyzed only after reintroduction into the wild as part of a rehabilitation program, 9 were analyzed both while in captivity or semi-captivity and after reintroduction, 20 were sampled only in captivity or semi-captivity, and 5 were free-living manatees. Serological tests were used to screen for antibodies against smooth Brucella spp. Whole blood, swabs, and tissue samples were screened for Brucella spp. DNA by PCR. Samples with positive PCR results were cultured for Brucella spp. isolation. All manatees yielded negative results in serological and molecular tests. Brucella spp. DNA was detected in the kidney of one adult Guiana dolphin Sotalia guianensis exhibiting necrosis in the liver. No growth of Brucella spp. was observed via microbiological culturing. This study is the first report of Brucella spp. DNA detection in cetaceans in the state of Pernambuco, and it highlights the importance of conducting systematic monitoring for the presence of Brucella infection in marine mammals along the Brazilian coast, especially in the northeast region, where several cases have been reported.

Long-term preservation of established fibroblast lines from six-banded armadillos (Euphractus sexcintus, Linnaeus, 1758) by extended passage and cryopreservation.

Establishing new somatic cell cultures has raised significant attention as an effective and convenient way to preserve genetic samples for different applications. Although many lines have been established in model animals, none derived from six-banded armadillo species is currently available. We report the successful isolation and characterization of fibroblasts from six-banded armadillos, evaluating the cell quality after extended culture and cryopreservation. Initially, we collected ear skin from five captive adult individuals and identified fibroblast lines by morphology, karyotyping, and immunophenotyping assays. The isolated fibroblasts were evaluated after several passages (fourth, seventh, and tenth passages) and cryopreservation by slow freezing. Cell morphology, viability, metabolism, proliferative activity, mitochondrial membrane potential, and apoptosis levels were analyzed. The skin explants had great adhesion, and cell outgrowth could be seen after 3-6 d. The cells were verified as fibroblasts at the fourth passage by vimentin expression and normal karyotype (2n = 58). The viability remained high (> 87%) and constant from the fourth to the tenth passage (p > 0.05). The passages did not change the cell morphology and metabolic and growth rates. Moreover, cryopreservation did not affect most evaluated parameters; post-thawed cells maintained their viability, growth, metabolism, and apoptosis levels. Nevertheless, cryopreservation increased mitochondrial membrane permeability and cell population doubling time compared to non-cryopreserved cells (p < 0.05). In summary, viable fibroblasts can be obtained from six-banded armadillo skin while conserving their quality as the number of passages increases and featuring few changes after cryopreservation.

Ex Situ Approaches for the Conservation of Genetic Resources in the Superorder Xenarthra.

Xenarthra-a superorder of placental mammals endemic to the Neotropics-is represented by armadillos, anteaters, and sloths. Considering their long history in the Americas, extant xenarthrans represent an important group for understanding the impact of past environmental changes on species diversification and serve key ecological functions as ecosystem engineers. Unfortunately, most wild xenarthran populations are at risk, due primarily to anthropogenic activities, necessitating urgent conservation efforts. Moreover, the paucity of information on some species has rendered population estimation and, consequently, conservation management challenging. In addition, relatively few groups are researching this superorder, perhaps because fieldwork with armadillos, anteaters, or sloths and their captive care are challenging tasks. Nevertheless, dedicated research and efforts to ensure the long-term conservation of these animals are deemed essential. In this context, cryobanks are a practical approach for breeding and maintaining genetic diversity in wildlife, and they are important tools for assisting and improving both ex situ and in situ conservation strategies. Therefore, cryopreservation of biological resources may be a promising strategy for conserving xenarthrans. Specifically, semen cryopreservation, which has already been applied in some species, may be the most effective strategy for this group. The present article provides an overview of ex situ conservation of xenarthrans, which will contribute to the development and implementation of additional strategies for protecting these unique mammals.

Viabilidade do sêmen de tatus-peba (Euphractus sexcinctus) centrifugado e diluído em Tris ou agua de coco em pó / Viability of the six-banded armadillos (Euphractus sexcinctus) semen centrifuged and extended in Tris or powdered coconut water

Foram avaliados os efeitos da centrifugação associada ao uso de dois diluentes na manutenção da viabilidade espermática em tatus-peba (Euphractus sexcinctus) ao longo do teste de termorresistência (TTR). Amostras de sêmen (n=12), oriundas de 04 machos adultos coletados por eletroejaculação, foram divididas em quatro alíquotas, sendo duas imediatamente diluídas em Tris ou água de coco em pó (ACP-119(r)), e as outras duas centrifugadas (800g10min-1) previamente à diluição. As amostras foram incubadas a 34°C por 3h, e os parâmetros seminais avaliados em intervalos de 1h. Em termos gerais, verificou-se uma redução da viscosidade espermática imediata à diluição em ambos os diluentes, independente do uso da centrifugação. Aos 60 minutos, verificou-se uma redução dos parâmetros avaliados (P<0,05), embora o Tris tenha promovido uma melhor preservação deles (P<0,05), quando comparado ao ACP-119(r) até os 120 minutos de avaliação. Após este período, os dois diluentes se equipararam (P>0,05). Ainda, verificou-se um efeito deletério da centrifugação sobre a qualidade do sêmen de tatus-peba durante todo o teste de termorresistência. Nas condições do presente estudo, conclui-se que o diluente Tris mostrou-se superior ao ACP-119(r) para a manutenção da viabilidade do sêmen de tatus-peba, sendo desnecessária a realização de centrifugação prévia à diluição.

The effects of the centrifugation associated to the use of two extenders on the viability of six-banded armadillo's (Euphractus sexcinctus) sperm were evaluated during a thermo resistance test (TRT). Semen samples (n=12) derived from 04 stud males collected by electroejaculation were divided in four aliquots; two of that were immediately diluted in Tris or powdered coconut water (ACP-119(r)); the two others were centrifuged (800g10min-1) prior to the dilution. Samples were incubated at 34ºC per 3h, and the semen parameters were evaluated at each hour. In general, dilution promoted a reduction in semen viscosity in the use of both diluents using centrifugation or not. At 60min, a decrease was verified for all semen parameters (P<0.05), however they were better preserved in the use of Tris when compared to ACP-119(r) (P<0.05) up to 120min. After that, both diluents equated. In addition, centrifugation procedure presented a deleterious effect on the armadillo's semen quality during all the thermoresistance test. In the present conditions, conclude that Tris extender is more efficient than ACP-119(r) for the preservation of six-banded armadillo semen viability, and the previous centrifugation is unnecessary.

Atividade antibacteriana e cicatrizante do óleo de buriti Mauritia flexuosa L. / Antibacterial and healing activities of buriti oil Mauritia flexuosa L.

O objetivo deste estudo foi avaliar a atividade antibacteriana in vitro e cicatrizante do óleo de buriti (M. flexuosa) em feridas realizadas em ratos (Rattus norvegicus albinus). Para a avaliação antibacteriana in vitro, foram utilizados cinco patógenos bacterianos incluindo espécies gram-positivas e espécies gram-negativas mediante o uso do método de difusão em ágar. Para a avaliação da atividade cicatrizante, foram utilizados 40 ratos da linhagem Wistar, divididos em dois grupos: o grupo I, composto por 20 ratos com feridas cutâneas, tratados com aplicação tópica do creme base com 10 por cento de óleo de buriti, e o grupo II, controle, com o mesmo número de animais que receberam a aplicação tópica do creme base. A aplicação do produto foi realizada em feridas padronizadas, circulares de 1cm de diâmetro na região dorsolombar. As avaliações clínica, morfométrica e histopatológica das feridas foram realizadas no 3°, 7°, 14° e 21° dias. Em relação à avaliação da atividade antibacteriana, os resultados mostraram que houve inibição do crescimento bacteriano em quatro dos cinco patógenos testados. Em relação à área da ferida, foi observada redução significativa da área no 14o dia e maior percentual de contração das feridas do grupo tratado em relação ao controle. No décimo quarto dia, as feridas tratadas com o óleo do buriti apresentavam aumento significativo na contagem de fibroblastos e fibras colágenas, além de completo processo de reepitelização, enquanto o grupo controle necessitava de mais tempo para resolução do processo cicatricial.

The aim of this study was to evaluate the in vitro antimicrobial activity and wound healing effect of buriti oil (M. flexuosa) in rats. To evaluate the in vitro antimicrobial activity, five species of bacteria, including both gram-negative and gram-positive, were tested by the agar diffusion method. To assess the wound healing effect, 40 rats of Wistar lineage were clustered into two groups: G1, composed by 20 rats with cutaneous wounds and treated using topic administration of basic cream containing 10 percent of buriti oil; and G2 or control group, composed by 20 rats with cutaneous wounds and treated using topic administration of basic cream without any buriti oil. The cream administration was performed on circular wounds of 1 cm area in the lumbodorsal region. Clinical, histopathologic and morphometric evaluations of the wounds were done in 3rd, 7th, 14th and 21th days. Four from five bacteria species tested had growing inhibition, which demonstrates the antimicrobial potential of buriti oil. A significant reduction on the wound area with contraction of the edges was found for G1 in the 14th day. On this same day, the wounds treated using buriti oil showed an increase in the fibroblasts and collagen fibers countings and complete reephitelialization, characteristics not demonstrated by G2.