Mango tree pruning hay in substitution of elephant grass in cattle diet

This experiment was conducted to evaluate the effect of different levels of substitution (0, 33.3, 66.7, and 100 %) of elephant grass by hay of mango tree pruning (HMTP) on intake and digestibility in cattle and on the in vitro gas production. Moreover, the effect of tannin in HMTP on the gas production and in vitro degradability of the dry matter was evaluated, using polyethylene glycol (PEG). The intake and digestibility were evaluated in a double 4?×?4 square Latin design, using eight Holstein intact bulls. To evaluate the gas production and degradability, in vitro semi-automated gas production technique was used. The substitution of elephant grass by HMTP decreased the intake (P?<?0.05) and the digestibility (P?<?0.05) of the nutrients and also the gas production (P?<?0.05). Furthermore, the utilization of PEG as a tannin-complexing agent increased the gas production and degradability of the HMTP (P?<?0.05). It was concluded that the high cell wall lignification and the presence of tannin limit the use of HMTP as the only source of roughage in cattle diets.     

Feline Endometrial Adenocarcinoma in Females <1 Year Old: A Description of Four Cases

Uterine neoplasms of epithelial origin are rare in cats and most often are described in older females. Yet, in less than 2 years, four ovariohysterectomy specimens were submitted from different practices to the Laboratory of Histology and Anatomical Pathology, at UTAD (Vila Real, Portugal), that emitted a diagnosis of feline endometrial adenocarcinoma. Untypically, all the females were aged <1 year old at the surgery. Access to the clinical files was requested to document the clinical features of the four cases, including any complementary data available, to construct the present case reports. The clinical situation developed with discrete signs, but vulvar discharge was present in three cases, ranging from bloody to brownish or colourless, and from purulent to mucous. The females were in dioestrus, although the oestrus remained unperceived in most cases. In this study, the four clinical situations are described and discussed on the basis of available literature, highlighting the aspects that may impair an early diagnosis and that may favour the progression of the disease and also that age should not be an excluding criteria when analysing the differential diagnosis list.     

Reproductive Development of Santa Inês Rams During the First Year of Life: Body and Testis Growth,Testosterone Concentrations,Sperm Parameters,Age at Puberty and Seminal Plasma Proteins

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two‐dimensional SDS‐PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 ± 0.7 to 54.3 ± 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non‐significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 ± 0.05 to 1.14 ± 0.24 × 10⁹ cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 ± 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 ± 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.     

Bovine ovarian stem cells differentiate into germ cells and oocyte‐like structures after culture in vitro

Stem cells have been isolated from ovaries, and their ability to differentiate into oocytes in vitro has been demonstrated for mice and human, but not for bovine species. The aims of this study were to isolate germline stem cells from bovine ovaries and to evaluate the effects of bone morphogenetic proteins (BMPs) 2 and 4, and follicular fluid on the differentiation of these stem cells into oocyte‐like structures. The ovarian stem cells were isolated and cultured in α‐MEM⁺ supplemented with BMP2, BMP4 or follicular fluid. On days 0 and 14, cells were evaluated for their morphological appearance, viability, expression of alkaline phosphatase and for markers of germ cell formation (VASA and DAZL) and oocyte development (GDF9, ZPA and SCP3) by qPCR. Levels of mRNA were analysed using ANOVA and Bonferroni test (p < .05). The results showed that at day 0, ovarian stem cells expressed specific markers of pluripotency (OCT4, SOX). In addition, these cells were positive for alkaline phosphatase, which is a marker commonly used to identify primordial germ cells (PGCs). After the period of differentiation, cells had morphological features that resemble PGCs and oocyte‐like cells (OLCs). An increase, ranging from five to 14 times, in the expression of VASA was observed in cells cultured in medium supplemented with BMPs and follicular fluid, while the increase in DAZL expression ranged from four to six times. In addition, OLCs had an increase in expression of mRNAs for GDF9, ZPA and SCP3 that ranged from two to eight times. In conclusion, OLCs can be differentiated in vitro from ovarian stem cells and BMPs and follicular fluid are effective in stimulating the expression of mRNAs for germ cell and oocyte markers.     

Interactions Among Different Devices and Electrical Stimulus on the Electroejaculation of Captive Agoutis (Dasyprocta leporina)

The interactions among different electroejaculation devices associated with serial or continuous stimuli were investigated to improve the efficiency of the electroejaculation for semen collection in agoutis. Ten sexually matured male Dasyprocta leporina were restrained by the intramuscular administration of xylazine–ketamine association. Each individual was randomly subjected to four electroejaculation protocols, by combining two devices (one presenting longitudinal electrodes emitting square waves and other presenting ring electrodes emitting sine waves) and two electrical stimuli protocols (serial or continuous). A total of 40 attempts for electroejaculation were conducted in agoutis, being 10 per treatment. The most efficient treatment in providing ejaculates containing sperm (p < 0.05) was that using and electroejaculator connected to a probe with ring electrodes and associated with serial stimuli (4/7; 57%). In spite of semen parameters obtained by sine waves were adequate for using the samples for assisted reproduction, higher values for sperm motility and functional membrane integrity were obtained in the use of the square wave, independently of the electric stimulation protocol used (p < 0.05). In conclusion, we verified that the use of a device presenting a probe with ring electrodes and emitting sine waves, associated with a serial stimuli protocol, improves the efficiency for semen obtaining by electroejaculation in adults D. leporina.     

Molecular diversity, genetic structure and mating system of Calopogonium mucunoides Desv.

Calopogonium mucunoides Desv. is a species native of South and Central America that is used as green manure and a pasture crop. The molecular genetic diversity was characterized in 195 C. mucunoides accessions from a germplasm collection using 17 microsatellite markers. Outcrossing rate was estimated after the evaluation of six microsatellite loci in 200 genotypes originated from 10 open-pollinated progenies (20 genotypes per progeny). Six genetic clusters were identified in the germplasm collection by the STRUCTURE software analysis, neighbor-joining tree comparisons and principal component analysis, which highly correlated with the geographic locations where these accessions were originated or collected. These results were confirmed using AMOVA. The largest portion of the genetic variation was observed among clusters (64.38%). The results indicated that: multilocus outcrossing rate (t _m ) was 16.3%, suggesting a mixed mating system with a predominance of autogamy; single locus outcrossing rate (t _s ) was 11%; difference (t _m ?t _s ) was 0.054, indicating that only 5.4% of outcrossing occurred among related individuals; paternity correlation (r _p ) was 33% suggesting a low probability of finding full sibs among the progeny; parental coefficient of inbreeding (F _m ) was 5.0%, indicating a low degree of inbreeding in each parent. A core collection for C. mucunoides was assembled to capture the allelic diversity found in this study. The complete allelic diversity was represented by only 15 accessions. These results should be useful for exploiting the genetic resources of C. mucunoides and could influence future conservation efforts and breeding programs.     

Intracellular reactive oxygen species production by polymorphonuclear leukocytes in bovine leukemia virus-infected dairy cows

The present study assesses the oxidative burst activity from polymorphonuclear leukocytes (PMNLs) from bovine leukemia virus (BLV)-infected cows. Fifteen clinically healthy cows were divided into serologically positive cows without any hematological alteration, serologically positive animals with persistent lymphocytosis (PL) and healthy serologically negative cows. The oxidative burst activity from the PMNLs was evaluated by flow cytometry using 2',7'-dichlorofluorescein diacetate as a probe. PMNLs from each cow were incubated with heat-killed Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) to stimulate oxidative burst activity. The results of the present work showed no significant difference in the oxidative burst activity without any stimulus and elicited by S. aureus. Conversely, a decrease in the oxidative burst index induced by E. coli in PMNLs was observed in BLV-infected cows.     

Pathogenic and molecular comparison of Puccinia kuehnii isolates and reactions of sugarcane varieties to orange rust

Sugarcane orange rust, a disease caused by Puccinia kuehnii, was first reported in Brazil in 2009. There are no studies comparing the Brazilian P. kuehnii collections and the reaction of important sugarcane varieties under controlled conditions. This work compared the reaction of seven sugarcane varieties inoculated with six different P. kuehnii isolates from Brazilian sugarcane areas and verified the pathogenic and genetic variability of these isolates. The incubation (I) and latency (L) disease periods, disease severity (SEV), total number of lesions (TNL), total number of sporulating lesions (TNSL), and percentage of sporulating lesions (%SL) were evaluated. Furthermore, ITS1 and IGS ribosomal sequences of all P. kuehnii isolates used in this study were compared with pathogen sequences from 13 different countries. The disease incubation ranged from 7 to 10 days and the latency ranged from 10 to 21 days. SEV and TNL showed large variations and few significant differences between the reaction of the varieties to P. kuehnii, in contrast with the variables TNSL and %SL. The P. kuehnii isolates did not compose different virulent races, but the isolate from one site (Araras) was a more aggressive race. The ITS1 and IGS ribosomal sequences of six P. kuehnii isolates were identical with each other and to most P. kuehnii American sequences deposited at GenBank. The studied sequences of P. kuehnii isolates differed from the sequences from Asia, Tahiti and Oceania.     

Native leguminous plants for mineland revegetation in the eastern Amazon: seed characteristics and germination

New Forests - For the effective use of native plants for mineland revegetation, an understanding of seed dormancy break and germination requirements, and seed storage tolerance is indispensable. In...