HPLC separation and determination of 12 cholesterol oxidation products in fish: comparative study of RI, UV, and APCI-MS detectors

A simple, fast, and sensitive method for the extraction through direct saponification, separation, quantification, and identification of 12 cholesterol oxidation products (COPs) and cholesterol in a single isocratic, normal-phase, high-performance liquid chromatography (HPLC) was developed. Three detectors were compared for determination of COPs and cholesterol in fish samples: refractive index (RI), ultraviolet (UV), and atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The results did not show significant differences (p > 0.05) between the concentration of the cholesterol oxides and cholesterol obtained with these detectors. The present study demonstrated the presence of 19-hydroxycholesterol, 22"R"-hydroxycholesterol, 22"S"-hydroxycholesterol, 24"S"-hydroxycholesterol, and 25"R"-hydroxycholesterol for the first time in fish samples.     

Mycobacterium bovis cultured from commercially pasteurized cows' milk: laboratory cross-contamination

The ability of Mycobacterium paratuberculosis to survive the commercial pasteurization process of raw milk remains controversial. In a study undertaken in Venezuela to culture M. paratuberculosis from commercially pasteurized cows' milk, 83-200 ml containers of milk were processed and cultured on Herrold's egg yolk slants. No M. paratuberculosis was cultured but a total of six colonies of Mycobacterium bovis were isolated from one container each from two different milk providers. Because laboratory cross-contamination was suspected, the laboratory records were reviewed and spoligotyping was carried out on the isolated individual colonies. On the day before these milk specimens were processed, the biological safety cabinet had been used for the isolation of M. bovis from lymph nodes from infected cattle. Spoligotyping showed that that the colonies isolated from the milk all had the same pattern as the strains isolated from the lymph nodes that were processed the previous day. As far as we know, this is the first report of cross-contamination in a veterinary mycobacterial laboratory. False-positive cultures in the mycobacterial laboratory are not rare. In this setting M. bovis was isolated because it is the most common manipulated organism in this laboratory. We believe that reports on the isolation of M. paratuberculosis from commercially pasteurized milk should exclude cross-contamination before reporting, especially when this organism is routinely isolated from animal material in the same lab.     

Cross-modality of texture and aroma perception is independent of orthonasal or retronasal stimulation

To assess the influence of orthonasal and retronasal stimulation on cross-modal interactions between texture and flavor perception of food, a series of experiments have been conducted. Healthy human subjects were exposed to strawberry aroma pulses delivered by a computer-controlled stimulator based on air dilution olfactometry. Just prior to exposure to the aroma, the human subjects consumed water, custard, or protein gels with different textures without any added aroma. The aroma was delivered as a sequence of aroma pulses, in either an orthonasal or a retronasal fashion. The retronasal presentation of aroma with concomitant presentation of texture is thought to more closely mimick the in vivo flavor release of semisolid food products as compared to orthonasal stimulation. The time between oral consumption of the food, including swallowing, and the exposure to the aroma varied between 0.5 and 6.5 s. The subjects rated the intensity of the strawberry aroma. It was observed that the intensity of aroma decreased with increasing firmness of the food that was consumed. Aroma pulses delivered 6.5 s after swallowing were perceived as being more intense as compared to aroma pulses delivered immediately after swallowing. In conjunction with late delivery, the effect cross-modal interactions apparently decreased. Significantly higher odor intensities were reported for the aroma stimuli supplied orthonasally in comparison to retronasal administration. The observed cross-modal effect of texture on aroma intensity was not significantly altered by the mode of aroma delivery, i.e., orthonasal or retronasal stimulus administration.     

Biological control of postharvest diseases of fruit

Postharvest decay in harvested fruit causes considerable economical losses. Fungicides are the primary means to control these losses. Public concern in food safety and environmental issues and the increase of pathogen resistant populations have enhanced the interest in developing alternative methods to fungicides to control postharvest fruit decay. During the last two decades a huge information and advances concerning the selection of antagonists, mode of action, different approaches to enhance biocontrol activity, formulation and production have been achieved, and some biofungicides are already in the market. It is likely that several more products will enter the market in the near future, as the result of the biological control research programs worldwide. Nonetheless, it is necessary to continue finding new potential microorganisms, better understanding the mode of action, and pathogen, antagonist and host interactions, to increase the potential of biocontrol helping to become a real alternative to synthetic postharvest fungicides. This article presents an overview of postharvest biological control approaches and explores new research possibilities to improve biocontrol activity.     

Fine-scale effects of habitat loss and fragmentation despite large-scale gene flow for some regionally declining woodland bird species

Habitat loss and associated fragmentation effects are well-recognised threats to biodiversity. Loss of functional connectivity (mobility, gene flow and demographic continuity) could result in population decline in altered habitat, because smaller, isolated populations are more vulnerable to extinction. We tested whether substantial habitat reduction plus fragmentation is associated with reduced gene flow in three ??decliner?? woodland-dependent bird species (eastern yellow robin, weebill and spotted pardalote) identified in earlier work to have declined disproportionately in heavily fragmented landscapes in the Box-Ironbark forest region in north-central Victoria, Australia. For these three decliners, and one ??tolerant?? species (striated pardalote), we compared patterns of genetic diversity, relatedness, effective population size, sex-ratios and genic (allele frequency) differentiation among landscapes of different total tree cover, identified population subdivision at the regional scale, and explored fine-scale genotypic (individual-based genetic signature) structure. Unexpectedly high genetic connectivity across the study region was detected for ??decliner?? and ??tolerant?? species. Power analysis simulations suggest that moderate reductions in gene flow should have been detectable. However, there was evidence of local negative effects of reduced habitat extent and structural connectivity: slightly lower effective population sizes, lower genetic diversity, higher within-site relatedness and altered sex-ratios (for weebill and eastern yellow robin) in 10 × 10?km ??landscapes?? with low vegetation cover. We conclude that reduced structural connectivity in the Box-Ironbark ecosystem may still allow sufficient gene flow to avoid the harmful effects of inbreeding in our study species. Although there may still be negative consequences of fragmentation for demographic connectivity, the high genetic connectivity of mobile bird species in this system suggests that reconnecting isolated habitat patches may be less important than increasing habitat extent and/or quality if these need to be traded off.     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

Evaluation of infertility in the bitch

The majority of bitches that are brought to a veterinarian for reproductive evaluation are actually healthy. Infertility in the bitch is most commonly due to inappropriate breeding management rather than an inability to conceive. Before embarking on an exhaustive medical evaluation, it is important to obtain a thorough history and physical examination to identify any problems. Completion of a routine database assists in detecting significant systemic illness that may impact infertility. After consideration of history, physical examination, and minimum database, fertility problems are most commonly considered in one of four categories: abnormal estrous cycles, normal estrous cycles, failure to breed, or failure to carry a litter to term. This classification system aids in construction of a differential list and systematic evaluation of all differentials. The majority of diagnostic tests required are available to the general practitioner. Before any treatment for infertility, adequate health management must be ensured. Specific treatment for the most common causes of infertility is centered on appropriate breeding management.     

The effect of body condition on propofol requirement in dogs

ObjectiveTo determine if body condition score (BCS) influences the sedative effect of intramuscular (IM) premedication or the dose of intravenous (IV) propofol required to achieve endotracheal intubation in dogs.Study designProspective clinical study.AnimalsForty–six client–owned dogs undergoing general anaesthesia.MethodsDogs were allocated to groups according to their BCS (BCS, 1 [emaciated] to 9 [obese]): Normal–weight Group (NG, n = 25) if BCS 4–5 or Over–weight Group (OG, n = 21) if BCS over 6. Dogs were scored for sedation prior to IM injection of medetomidine (5 μg kg^?1) and butorphanol (0.2 mg kg^?1) and twenty minutes later anaesthesia was induced by a slow infusion of propofol at 1.5 mg kg^?1 minute^?1 until endotracheal intubation could be achieved. The total dose of propofol administered was recorded. Data were tested for normality then analyzed using Student t–tests, Mann–Whitney U tests, chi–square tests or linear regression as appropriate.ResultsMean ( ± SD) propofol requirement in NG was 2.24 ± 0.53 mg kg^?1 and in OG was 1.83 ± 0.36 mg kg^?1. The difference between the groups was statistically significant (p = 0.005). The degree of sedation was not different between the groups (p = 0.7). Post–induction apnoea occurred in 11 of 25 animals in the NG and three of 21 in OG (p = 0.052).ConclusionsOverweight dogs required a lower IV propofol dose per kg of total body mass to allow tracheal intubation than did normal body condition score animals suggesting that IV anaesthetic doses should be calculated according to lean body mass. The lower dose per kg of total body mass may have resulted in less post–induction apnoea in overweight/obese dogs. The effect of IM premedication was not significantly affected by the BCS.Clinical relevanceInduction of general anaesthesia with propofol in overweight dogs may be expected at lower doses than normal–weight animals.     

Improving efficiency of sow productivity： nutrition and health

This reviews research focused to understand the nutrient requirement and balance to meet the needs of fetal growth,mammary growth,and milk production.This summary will handle how feeding strategies can be adjusted according to the nutrient needs for a sow to enhance productivity and health.Most research data used in this summary are based on the studies conducted by the authors between 1996 and 2013.Nutrient requirements of sows are affected by stage of gestation and parity of sows.Dietary antioxidant concentrations need to be re-evaluated for its sufficiency in sow diets especially to prevent excessive oxidative stress during late gestation and lactation.When feeding sows,consideration of phase feeding of gestating sows and parity feeding of lactating sows could enhances production longevity and health of sows.Use of selected nutrients and additives seems to help productivity and health of sows.