The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Protein requirements of collared peccary (<Emphasis Type="Italic">Pecari tajacu</Emphasis>)

A nitrogen (N) balance digestion trial was conducted to determine the protein requirement of collared peccaries (Pecari tajacu). In a 4 × 4 Latin square design, four captive adult male peccaries were fed four isoenergy diets containing four different levels of N (11.7, 16.3, 22.8, and 26.7 g N/kg of dry matter—DM). After 15 days of adaptation, a total collection of feces and urine was carried out for five consecutive days. Regression analyses between N intake and N in feces and urine allowed to calculate the metabolic fecal nitrogen (MFN = 2.3 g N/kg of dry matter intake—DMI) and daily endogenous urinary N (EUN = 185 mg N/kg^0.75). Likewise, by regression analyses between consumption of nitrogen and the nitrogen balance (NB = N ingested ? N excreted, mg N/kg^0.75), a daily requirement of 514 mg N/kg^0.75 was calculated. Therefore, if food intake is unrestricted, collared peccaries require a minimum in their diet of about 5.4% crude protein on DM basis. These values are almost as low as those found for browsing and frugivorous wild ruminants, which reinforce the proposition that peccaries’ digestive physiology is nearer to that of domestic and wild ruminants than domestic pigs. This relatively low protein requirement of collared peccary and its great ability to digest protein reveal the relevance of the forestomach for the species on nitrogen/protein metabolism and allow the use of diets with lower crude protein levels than the commercial ones used for the domestic pig, which reduces feed costs.     

Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts.

Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum.

Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used.

Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ～10⁰ parasite mL^?1. For the blood samples, 40/69 (58%; CI 95%; 46%–69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing.

Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.     

Can facultative parthenogenesis occur in biparental mealybug species?

The existence of facultative parthenogenesis in mealybugs has been reported by different authors and questioned by others in relation to certain species. We tested the hypothesis of facultative parthenogenesis in relation to three mealybug species, Planococcus citri (Risso), Pseudococcus calceolariae (Maskell) and Pseudococcus viburni (Signoret). The results suggest that these mealybug species are obligate amphimictic.     

Intraspecific variation in 16S rRNA gene of Mycoplasma synoviae determined by DNA sequencing

Mycoplasma synoviae (MS) is an important avian pathogen may cause both respiratory disease and joint inflammation synovitis in poultry, causing economic losses to the Brazilian poultry industry. The genotypic variation in 16S rRNA gene is unknown. Partial sequences of 16S rRNA gene of 19 strains of M. synoviae were sequenced and analyzed in order to obtain molecular characterization and evaluation of the genetic variability of strains from distinct Brazilian areas of poultry production. Different polymorphic patterns were observed. The number of polymorphic alterations in the studied strains ranged from 0 to 6. The nucleotide variations, including deletion, insertion and substitutions, ranged from 3 to 5. The genotypic diversity observed in this study may be explained by spontaneous mutations that may occur when a lineage remains in the same flock for long periods. The culling and reposition in poultry flocks may be responsible for the entry of new strains in different areas.     

Nutritional evaluation of “Braquiarão” grass in association with “Aroeira” trees in a silvopastoral system

This study aimed at evaluating the influence of a silvopastoral system (SPS) composed of “Aroeira” trees (Myracrodruon urundeuva) on the nutritional value of Brachiaria brizantha cv. Marandu forage grass over the rainy season. The experiment was conducted in the SPS located in the “Cerrado” biome in Lagoa Santa County, Minas Gerais state, Brazil, in the agricultural year of 2004–2005. It was evaluated the influence of “Aroeira”, whose density in the area was 140 trees/ha, on the following parameters of the B. brizantha cv. Marandu forage grass: forage mass harvest above stubble height, morphogenic attributes, bromatological composition and nutritional value. The effects of “Aroeira” trees on the micro-climatic conditions were also evaluated. The experiment adopted a randomized block design with five blocks (cutting seasons), two treatments (a shaded pasture—B. brizantha cv. Marandu shaded by M. urundeuva—and a pasture in full sun—B. brizantha cv. Marandu in full sun), and three replicates. The tree component, composed of the tree species M. urundeuva in the experimental density adopted, increased dry matter and crude protein of the forage mass harvest above stubble height, without affecting the concentration of neutral detergent fibre or acid detergent fibre, thus causing dry matter and organic matter in vitro degradability as well as effective degradability of the forage grass to remain the same as those obtained in full sun. Therefore, the results of this study show the possibility of using trees and grass to increase both animal production and environmental services.     

Different approaches for modeling grouped survival data: A mango tree study

Interval-censored survival data, in which the event of interest is not observed exactly but is only known to occur within some time interval, occur very frequently. In some situations, event times might be censored into different, possibly overlapping intervals of variable widths; however, in other situations, information is available for all units at the same observed visit time. In the latter cases, interval-censored data are termed grouped survival data. Here we present alternative approaches for analyzing intervalcensored data. We illustrate these techniques using a survival data set involving mango tree lifetimes. This study is an example of grouped survival data.     

Chronic toxicity of fenoxycarb to the midge Chironomus riparius after exposure in sediments of different composition

Background, aim, and scope  

Fenoxycarb is an insect growth regulator widely used to control a variety of pests. As a juvenile hormone analogue, it interacts with the natural hormones involved in insect development, inhibiting metamorphosis to the adult stage. Adverse effects of fenoxycarb on non-target organisms have been repeatedly observed and its rapid dissipation from water to the organic fraction of the sediment is known. The aim of this work was to investigate the impact of fenoxycarb on the development of sediment-dwelling larvae of the midge Chironomus riparius after exposure to spiked sediment.     

Perturbation of the EphA2-EphrinA1 System in Human Prostate Cancer Cells by Colonic (Poly)phenol Catabolites

The Eph tyrosine kinase receptors and their ephrin ligands play a central role in human cancer as their deregulated expression induces tumorigenesis with aggressive phenotypes. To evaluate their potential contribution to EphA2-ephrinA1 modulation, several colonic catabolites of dietary (poly)phenolics, known to be generated in vivo, were screened using an ELISA-based binding assay. Some of the catabolites inhibited the binding in a dose-dependent manner (IC(50) values from 0.26 to 43 μM). Functional studies on prostate adenocarcinoma cells revealed that pyrogallol and protocatechuic acid specifically antagonized ephrinA1-Fc-induced EphA2 phosphorylation at concentrations that were not cytotoxic. The active concentrations of pyrogallol appear to be close to what can be reached in vivo under physiological conditions. Finally, because of the roles played by the Eph-ephrin system not only in cancer development but also in neurodegeneration and diabetes, pyrogallol and protocatechuic acid are candidates for more detailed functional studies to elucidate their role in these pathophysiological processes.