Cinthia Távora de Albuquerque Lopes

Multifocal Cutaneous Orf Virus Infection in Goats in the Amazon Region, Brazil

Orf virus is the etiological agent of contagious ecthyma, a severe exanthematic disease that affects small ruminants. Orf virus is zoonosis that is associated with occupational contact with infected animals in human disease. Clinically, contagious ecthyma is characterized by the appearance of vesicles, pustules, ulcers, and papillomatous proliferative lesions on the skin of the lips and nostrils. Here we describe a case of lethal cutaneous multifocal Orf virus infection in goats in the Amazon region of Brazil. Exanthematic lesions were collected and epidemiological and clinical data were obtained. Orf virus was detected using PCR amplification of the whole B2L, VIR, and VEGF open reading frame. Phylogenetic analysis revealed that this virus clustered together with the Orf virus samples isolated during classical contagious ecthyma. The present work is the first to report a severe proliferative Orf virus case in South America.

Prevalência sorológica e molecular de Babesia bovis e Babesia bigemina em búfalos (Bubalus bubalis) na Ilha de Marajó, Pará

O objetivo do estudo foi testar a prevalencia sorologica e molecular de Babesia bovis e Babesia bigemina em bufalos da Ilha de Marajo, Para. Foi utilizado ensaio de imunoadsorcao enzimatico indireto (iELISA) com antigeno total contendo proteinas de superficie externa e reacao em cadeia da polimerase (qPCR), envolvendo o uso de SYBR Green com base na amplificacao de um pequeno fragmento de gene do citocromo b. A prevalencia de animais positivos no ELISA para B. bovis, B. bigemina e para infeccao mista foi de 24.87% (199/800), 20.75% (166/800) e 18.75% (150/800), respectivamente. Na PCR foi detectado a presenca de B. bovis em 15% (18/199) e de B. bigemina em 16% (19/199) dos animais, sendo que destes, 58% (11/19) apresentavam-se co-infectados pelos dois agentes. Os resultados mostram uma baixa prevalencia de anticorpos anti-B. bovis e anti-B. bigemina em bufalos da Ilha do Marajo. Porem, observou-se que os agentes da babesiose bovina circulam em bufalos, podendo estes atuar como reservatorios.

Intoxicação por Cestrum laevigatum (Solanaceae) em bubalinos

Based on the history and clinical and pathological data, as well as on inspection of the pastures, a mortality of buffaloes in the county of Itaguai/RJ, Brazil, was diagnosed as poisoning by Cestrum laevigatum Schlecht., a plant of the Solanaceae family. The poisoning was reproduced in two buffaloes. Dried leaves of the shrub were administered by hand, in single doses corresponding to 20g/kg and 40g/kg of the fresh leaves, to four buffaloes of the Murrah breed. The dose corresponding to 40g/kg of the fresh leaves caused signs of poisoning, as apathy, anorexia, absence of rumen movements, dysmetria, excitement and aggressiveness, followed by death of the two buffaloes within 65 hours after administration. From the two buffaloes that received the corresponding dose of 20g/kg of the fresh plant, one presented clinical signs characterized mainly by decrease of the rumen movements, but recovered 97h22min after the administration; the other buffalo did not show symptoms of poisoning. Laboratory analyses for biochemical evaluation accused hepatic alterations. In one buffalo that died, the main macroscopic finding was an orange liver with a clear nutmeg appearance; in the second buffalo, the orange liver had no nutmeg appearance. Other alterations found in these two buffaloes were slight edema of the gall bladder wall, a slightly reddish mucous membrane of the abomasum, extensive echymoses in the endocard of the left ventricle and few petechiae in the endocard of the right ventricle; the abomasum content was slightly dry, and the large intestine had little and slightly dry contents wrapped by mucus. Histopatological examination revealed severe coagulative necrosis of the liver parenchyma in the centrolobular and intermediate lobular areas, with a halo of vacuolated hepatocytes at the periphery of the necrotic areas.

Quadro clínico-patológico do envenenamento crotálico experimental em bubalinos comparado com o de bovinos

The objective of the study was to verify the sensibility of buffaloes to the poison of Crotalus durissus terriiicus and to study the clinical-pathological picture in buffaloes in comparison with the one in cattle. The subcutaneous inoculation of the liofilized poison of the snake, diluted in 1ml of physiologic solution, was done in the area of the humerus-radio-ulnar joint of three buffaloes at doses of 0.015, 0.03 and 0.066mg/kg, and of two cattle at doses of 0.03 and 0.066mg/kg. The buffalo that received the 0.03mg/kg dose presented severe clinical signs but recovered six days later. The bovine that received the same dose, died after a clinical course of 22h56min. The 0.066mg/kg dose caused death of the bovine as also the buffalo, with a clinical course of 4h23min and 8h12min, respectively. The buffalo that received the 0.015mg/kg dose recovered, after a course of 48 hours. The buffalo that died, showed clinical signs from 3h58min on, and the buffaloes that showed symptoms from 17h25min and 24h00min after inoculation of the venom, but recovered. In the two cattle that died (with doses of 0.03 and 0.066mg/kg), the first clinical signs occurred 6h10min and 6h31min after the inoculation of the venom. The inoculation produced in the buffaloes and cattle nervous symptoms of flaccid paralysis. The main signs observed in the buffaloes as well as the cattle, were slight volume increase at the site of inoculation, respiration difficulties characterized by predominantly abdominal breathing, apathy, sialorreia, difficulty to get up when stimulated, evolution to sternal decubitus followed by lateral decubitus with peddling movements of the legs, and decrease of the reflexes related to the cranial nerves. The buffaloes showed also augmentation of the sustentation base, dragging of the hooves of the hind legs, slow and staggering gait, difficulty in apprehension of the food, The cattle showed additionally paralysis of the eyeballs, revealed through non-exhibition of the sclera during head rotation in latero-caudal direction. Laboratory exams revealed in the cattle and the buffaloes leucocytosis by neutrofilia, and in the biochemistry series, increase in the levels of alanine aminotransferase, aspartato aminotransferase, creatinaquinase and lactic dehydrogenase. There was no alteration in the urinalysis nor in the activation time of protrombine and in the time of partially activated tromboplastin. Necropsy only evidenced slight edema corresponding to the inoculation site in one bovine. Histopathological examination revealed picnosis of the epitelial cell nuclei of some kidney tubules in the cortex (in the buffalo and in one bovine) and slight vacuolation of hepatocites (in one bovine).

Proliferation of human adipose tissue-derived stem cells stimulated by oil rich in thymol of Lippia origanoides

PURPOSE To evaluate the effects of this thymol-rich oil in the proliferation of human adipose tissue-derived stem cells. METHODS Stem cells were isolated from human adipose tissue by liposuction. After the first passage, cells were cultivated in triplicate for three days in control medium and medium supplemented with three oil samples (1.0 μg/mL, 5.0 μg/mL, and 25.0 μg/mL). Cells were analyzed by the MTT assay at passage 1 (P1), and cell proliferation of control and 1 μg/mL groups was determined with a hemocytometer at P2 and P3. RESULTS Viability of the essential oil-treated cells was significantly higher than the control group at P1 (p = 0.0008). The treatment with the oil, at a concentration of 1 µg/mL, led to increases of 24.8% at P1 and 43.0% at P3 in the rate of cell proliferation compared with control cells. CONCLUSION Supplementing culture medium with essential oil of Lippia origanoides increased cell proliferation, especially at later passages.

Vitrification of Ovarian Tissue from Non-Human Primates

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue. Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened primate species and the needs to preserve their habitat, but also their gametes, development of preservation protocols are needed. Among the procedures, vitrification appears as a practical method to be applied in the near future. Although a low number of studies is reported, most of them were performed in the recent years. In this context, this article reviews recent information on the vitrification of ovarian tissue of non-human primates. Due to the limited number of studies in these species, observed data are compared with the literature in domestic and human mammals. Conclusion: Despite the advances summarized here, it is possible to conclude that there is still a need to improve procedures for the preservation of genetic material, including in post-mortem situations. The technique of vitrification, therefore, consists of a feasible and promising option. However, it is needed to study properly the methods of vitrification, as well as the introduction of new systems and supplements to minimize or avoid the cell damage caused by the cryopreservation procedure (physical and chemical).

Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells

PurposeDespite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development.Materials and methodsIn experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 103 and 104 cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control). In experiment 2, co-culture with 104 b-ATMSCs/mL was compared to the traditional granulosa cell co-culture system (Gran).ResultsIn experiment 1, co-culture with 104 b-ATMSCs/mL improved blastocyst rates in comparison to conditioned and control media (p < 0.05). Despite that it did not show difference with 103 b-ATMSCs/mL (p = 0.051), group 104 b-ATMSCs/mL yielded higher results of blastocyst production. In experiment 2, when compared to group Gran, co-culture with 104 b-ATMSCs/mL improved not only blastocyst rates but also quality as assessed by increased total cell numbers and mRNA expression levels for POU5F1 and G6PDH (p < 0.05).ConclusionsCo-culture of bovine embryos with b-ATMSCs was more beneficial than the traditional co-culture system with granulosa cells. We speculate that the microenvironmental modulatory potential of MSCs, by means of soluble substances and exosome secretions, could be responsible for the positive effects observed. Further experiments must be done to evaluate if this beneficial effect in vitro also translates to an increase in offspring following embryo transfer. Moreover, this study provides an interesting platform to study the basic requirements during preimplantation embryo development, which, in turn, may aid the improvement of embryo culture protocols in bovine and other species.

Detecção sorológica e molecular de Anaplasma marginale em búfalos na Ilha de Marajó, Pará

The aim of the study was to test the molecular and serological prevalence of Anaplasma marginale in water buffaloes of the Marajo Island, State of Para, Brazil. For serologic research were randomly selected 800 buffaloes and for molecular research 50 of these animals were randomly chosen. To quantify the serological prevalence we used the indirect enzyme linked immunosorbent assay (iELISA) with total antigen containing proteins outer surface. To quantify the prevalence molecular was used the polymerase chain reaction (PCR) involving gene amplification fragment larger surface protein 5 (MSP5). The prevalence of positive animals in iELISA was 25% (200/800). In the PCR we detected the presence of A. marginale in 2% (1/50) of animals. Although only one animal was positive in PCR, we found that it was negative in ELISA. The presence of the agent, even in low prevalence, shows that buffaloes can act as an important reservoir for transmission of the pathogen to cattle in northern Brazil.