Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Total vertebrectomy for stabilisation of chronic spinal lumbar luxation in a paraplegic dog without nociception

An adult male crossbred dog was referred with a history of a road traffic accident that took place 1 month earlier. Neurological examination revealed paraplegia with absent nociception in the pelvic limbs. On epaxial palpation, significant curvature of the anatomical axis of the spine between the third and fourth lumbar vertebrae was observed, with the presence of a bone end almost piercing the dog's skin. Survey radiographs of the lumbar spine revealed severe dislocation between L3 and L4 vertebrae. During surgery, the spinal cord was not visible between the dislocated segments. Because of difficulties in reducing the lumbar luxation during surgery, vertebrectomy and vertebral shortening were performed. After alignment between vertebrae L3 and L5, eight cortical orthopaedic screws and bone cement were used for fixation. After 30 days, the dog started to use a wheelchair and was considered by its owner to have a good quality of life with no evidence of pain. To the authors’ knowledge, this is the first case of severe luxation treated by total vertebrectomy and spine shortening in a dog. This surgery can be considered as an option in the management of severe spine luxation when the spinal cord is physically transected.     

Effects of progestagen exposure duration on estrus synchronization and conception rates of crossbreed ewes undergoing fixed time artificial insemination

Synchronization of estrus and ovulation are of paramount importance in modern livestock improvement programs. These methods are critical for assisted reproduction technologies, including artificial insemination and embryo transfer, that can increase productivity. In the current study, subcutaneous implants containing norgestomet were placed for long (14 days), medium (9 days), and short (5 days) periods of time in 70 crossbred ewes undergoing fixed-time artificial insemination. The resulting effects on estrus synchronization and conception rates were subsequently evaluated. Among the synchronized ewes, 85.7% (60/70) underwent estrus over a period of 72 h after progestagen treatment ceased. The shortest mean interval between withdrawal of the device and onset of estrus (34.2 ± 8.9 h) was observed in the G14 days of P4 group (p < 0.05). The conception rate of the G14 days of P4 group was statistically higher than that of the other groups (83.3% vs. 60.9% vs. 47.8%; p < 0.05). In conclusion, 14 days of norgestomet treatment produced higher conception rates and a greater number of pregnancies at the beginning of the breeding season.     

Inheritance of the stay-green trait in tropical maize

Stay-green maize genotypes have been associated with tolerance to biotic and abiotic stresses, including tolerance to drought, and to stalk and root lodging, but there is limited information on its inheritance. Thus, this research was conducted to study the inheritance of the stay-green trait using both conventional analysis and QTL mapping of the Design III in a tropical maize population developed from two inbred lines genetically divergent for this trait. Two-hundred and fifty F₂ plants were genotyped with 177 microsatellite markers, and their backcrossed progenies to both parental inbreds were evaluated at three locations. Ten plants per plot were assessed 120 days after sowing and the plot means scores for stay-green, adjusted for days to silking emergence, were used for analysis. The additive variance was larger than the dominance variance, the genetic by location interaction variance presented a high magnitude, and the heritability coefficient on a plant-basis a low magnitude. Seventeen QTL were mapped, most of them were clustered on four chromosomes and accounted for by 73.08 % of the genetic variance. About half of the QTL interacted with location, and the average level of dominance was partial dominance. The additive effects were larger than the dominance effects; the latter were not unidirectional, so that heterosis could not be exploited in crosses. Procedures for marker-assisted selection to increase the level of stay-green are discussed and an approach is suggested for using both stable and non-stable QTL in a marker-assisted backcross program.     

Aspects on Hormonal Control of Normal and Induced Parturition in the Dog

Average length of gestation on the dog is 63 ± 2 days but may vary between 57-71 days due to the long period of receptivity at oestrus and the extended period of sperm survival in the female genital tract. In contrast to other domestic animals progesterone- and oestrogen concentrations are almost identical in pregnant and non pregnant bitches, except for their rapid decline immediately prior to parturition. Control of luteolysis still poorly understood. Experiments with indomethacin leading to a blockade of the prepartal PgF_2α increase, which commences with the decrease of progesterone, point toward a role of PgF_2α at this stage of pregnancy, which was extended for several days. At physiological conditions first visible signs of parturition were observed at peak-PgF_2α levels, 33.6 ± 17.6 hours after onset of luteolysis, which lasted over 16.8 ± 3.4 hours. Pulse-type releases of oxytocin were only observed after this point of time. To test for the effect of progesterone-withdrawal, four 51-57 days pregnant bitches were treated with the antiprogestin RU 38486 which inhibits the activity of progesterone at the receptor level. In all dogs first visible signs of parturition were observed 33.5 ± 7.5 hours after onset of treatment. However, the process of parturition came to an end after cervical opening and totally only one puppy was born. Different to a normal parturition no increase of PgF_2α was observed. Relaxin levels were not influenced by treatment. These observations suggested that treatment with antiprogestin followed by PgF_2α might be an adequate method to induce parturition in the dog; first experiences seem to confirm this conclusion.     

Effect of the addition of Chaetoceros calcitrans,Navicula sp. and Phaeodactylum tricornutum (diatoms) on phytoplankton composition and growth of Litopenaeus vannamei (Boone) postlarvae reared in a biofloc system

The aim of this study was to evaluate the effect of the addition of Chaetoceros calcitrans, Navicula sp. and Phaeodactylum tricornutum (diatoms) on phytoplankton composition and the growth of Litopenaeus vannamei postlarvae reared in a biofloc system . Four treatments were used: BFT (biofloc system without feed and no addition of diatoms); BFT‐F (biofloc system with feed and no addition of diatoms); BFT‐D (biofloc system with the addition of diatoms and no feed); and BFT‐FD (biofloc system with the addition of feed and diatoms), all in triplicate. The shrimp (16 ± 0.02 mg) were stocked at 2500 postlarvae m^?3 and fed a commercial feed. Diatoms were added on the 1st, 5th, 10th and 15th day at a density of 5 × 10⁴ cells mL^?1 for each species. No significant differences (P > 0.05) between treatments were observed for gross primary production, net ecosystem production and water column respiration rate. However, significant differences (P < 0.05) were observed for nitrite, orthophosphate, alkalinity, final weight, weight gain, yield, feed conversion ratio (FCR), phytoplankton and cyanobacteria composition. The BFT‐FD treatment had better performance parameters for final weight (270 mg), weight gain (254 mg), yield (0.67 Kg m^?3) and FCR (0.61), indicating the benefits of the diatoms C. calcitrans, Navicula sp. and P. tricornutum for decreasing cyanobacteria and improving growth of L. vannamei postlarvae reared in biofloc systems.     

Lipids and Composition of Fatty Acids of Saccharina latissima Cultivated Year-Round in Integrated Multi-Trophic Aquaculture

This study is evaluating the seasonal lipid and fatty acid composition of the brown seaweed Saccharina latissima. Biomass was sampled throughout the year (bi-monthly) at the commercial cultivation site near a fish farm in an integrated multi-trophic aquaculture (IMTA) and at a reference site in Denmark (2013–2014). Generally, there was no difference in the biomass composition between sites; however, significant seasonal changes were found. The lipid concentration varied from 0.62%–0.88% dry weight (DW) in July to 3.33%–3.35% DW in November (p < 0.05) in both sites. The fatty acid composition in January was significantly different from all the other sampling months. The dissimilarities were mainly explained by changes in the relative abundance of 20:5n-3 (13.12%–33.35%), 14:0 (11.07%–29.37%) and 18:1n-9 (10.15%–16.94%). Polyunsaturated fatty acids (PUFA’s) made up more than half of the fatty acids with a maximum in July (52.3%–54.0% fatty acid methyl esters; FAME). This including the most appreciated health beneficial PUFA’s, eicosapentaenoic (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), but also arachidonic (ARA) and stearidonic acid (SDA), which are not found in land vegetables such as cabbage and lettuce. Compared to fat (salmon) and lean fish (cod) this seaweed species contains higher proportions of ARA and SDA, but lower EPA (only cod) and DHA. Conclusively, the season of harvest is important for the choice of lipid quantity and quality, but the marine vegetables provide better sources of EPA, DHA and long-chain (LC)-PUFA’s in general compared to traditional vegetables.     

Molecular detection of Leishmania sp. in cats (Felis catus) from Andradina Municipality, São Paulo State, Brazil

The aim of this work was to molecularly detect Leishmania species in 52 cats from Andradina Municipality, S?o Paulo State, Brazil. The direct parasitological test was performed by using imprints of poplited lymph node, bone marrow and spleen to verify amastigote forms of Leishmania spp. The samples that were positive parasitological tests were subjected to molecular analysis (PCR) and sequencing. Infection was detected for 5.76% (3/52) of the examined cats and two had presence of amastigote forms of Leishmania spp. in lymph nodes. Polymerase chain reaction (PCR) of kinetoplast minicircle DNA, indicated positive amplification for samples of spleen and lymph nodes and the sequencing resulted in 97% similarity with Leishmania (L.) chagasi. This study proved the occurrence of infection with Leishmania (L.) chagasi in felines from Andradina municipality, S?o Paulo State.     

Leishmania chagasi infection in cats with dermatologic lesions from an endemic area of visceral leishmaniosis in Brazil

Although dogs are considered the main domestic reservoirs for Visceral Leishmaniosis (VL), which is caused in the Americas by Leishmania chagasi, infected cats have also been recently found in endemic areas of several countries and became a public health concern. Accordingly, the purpose of this study was to evaluate cats with dermatologic lesions from an endemic area of VL and the natural infection of L. chagasi. A total of 55 cats were selected between April 2008 and November 2009 from two major animal shelters of Ara?atuba, Southeastern Brazil. All cats underwent general and dermatologic examinations, followed by direct parasitological examination of lymphoid organs, immunosorbent assay (ELISA) and indirect immunofluorescence (IFAT). In addition, detection of amastigotes was performed by immunohistochemistry (IHC) in skin lesions of all cats. VL was diagnosed in 27/55 (49.1%) cats with dermatological problems. Amastigotes were found in lymphoid organs of 10/27 (37.0%) cats; serology of 14/27 (51.9%), 6/27 (22.2%) and 5/27 (18.5%) cats was positive for ELISA, IFAT and both, respectively. The IHC identified 9/27 (33.3%) cats; 5/27 (18.5%) were positive only for IHC and therefore increased the overall sensitivity. Specific FIV antibodies were found in 6/55 (10.9%) cats, of which 5/6 (83.3%) had leishmaniosis. Real time PCR followed by amplicon sequencing successfully confirmed L. chagasi infection. In conclusion, dermatological lesions in cats from endemic areas was highly associated to visceral leishmaniosis, and therefore skin IHC and differential diagnosis of LV should be always conducted in dermatological patients in such areas.