Grafts of porcine small intestinal submucosa seeded with cultured homologous smooth muscle cells for bladder repair in dogs

Background

Due to numerous complications associated to gastrointestinal augmented cystoplasty, this study aimed to analyze the anatomic repair of the bladder of 10 female dogs using grafts of porcine small intestinal submucosa (SIS) seeded with cultured homologous smooth muscle cells, and compare them with the acellular SIS grafts.

Results

We assessed the possible side effects and complications of each type of graft by clinical examination, abdominal ultrasound and laboratory findings. Anatomic repair of neoformed bladder was assessed by histological staining for H/E and Masson''s Trichrome, analyzed with a Nikon Photomicroscope connected to the system of image analysis Image J.

Conclusions

We propose that SIS associated to homologous smooth cells can improve the quality of tissue repair, and consequently decrease the potential complications inherent to acellular SIS.     

Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

A size-based stock assessment model for invasive blue catfish in a Chesapeake Bay sub-estuary during 2001–2016

Stock assessment modeling provides a means to estimate the population dynamics of invasive fishes and may do so despite data limitations. Blue catfish (Ictalurus furcatus) were introduced to the Chesapeake Bay watershed to support recreational fisheries but also consume species of conservation need and economic importance. To assess management tradeoffs, managers need to understand the current status of the population and anticipate future population abundance and trends. A Bayesian size-based stock assessment model was used to estimate blue catfish abundance, fishing mortality, and size structure over time (2001–2016) in the tidal James River. The model estimated population size increases until around 2006, with declines in total abundance after 2011 and large blue catfish (≥80 cm total length) after 2001. These first estimates of blue catfish population dynamics in the Chesapeake Bay region provide inputs for projection models to evaluate prospective management actions and identify monitoring needs.     

Assessment of microbial biocontrol agent (BCA) viability to mechanical and thermal stress by simulating spray application conditions

Background

In order to improve the biological control agent (BCA) efficacy, stress factors threatening the viability of microorganisms during spray application need to be determined. The effect of spray mixture temperature and exposure time on Trichoderma harzianum T 22 and Bacillus amyloliquefaciens QST713 viability were tested. Concurrently the combined effect of mechanical and thermal stress effect on BCA viability were tested at two initial spray mixture temperatures (14 and 25 °C) by simulating a spray application using airblast sprayers featured by different tank capacity and a spray liquid circuit (without and with hydraulic agitation system). To assess the BCA microorganism viability, spray mixture samples were collected at time intervals along trials and plated to count the colony forming units (CFU).

Results

The critical temperature threshold that inhibited BCA viability was 35 °C with 30 min of exposure. The sprayer type, the initial temperature of the spray mixture and the temperature increment during the trials significantly decreased the number of CFU recovered. When simulating a spray application, the spray mixture temperature increase rate was determined mainly by the residual amount of spray mixture in the tank. Even if the tank capacity does not substantially affect the final temperature reached by the spray mixture, the higher residual spray mixture in bigger tanks can expose the BCAs for a longer time to critical temperatures.

Conclusions

Experimental trials allowed us to identify the effect of factors affecting the viability of tested BCAs, providing information about the actual chance to guarantee the biological efficacy of BCA treatments. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Multiplex real-time PCR SYBR Green for detection and typing of group III Clostridium botulinum

Clostridium botulinum type C and type D belonging to the group III organisms, are mainly responsible for animal botulism outbreaks. Clinical signs alone are often insufficient to make a diagnosis of botulism and a laboratory confirmation is required. Laboratory confirmation can be performed by demonstrating the presence of botulinum neurotoxins in serum, gastrointestinal contents, liver, wound of sick or dead animals, or by demonstrating the presence of C. botulinum in gastrointestinal contents, liver, and wound. Demonstration of spores in gastrointestinal contents or tissue of animals with clinical signs indicative of botulism reinforces the clinical diagnosis. With the aim of detecting and typing C. botulinum group III organisms, a multiplex real-time PCR SYBR Green was developed and in-house validated. Selectivity, limit of detection, relative accuracy, relative specificity, relative sensitivity, and repeatability of the method were investigated. The multiplex real-time PCR SYBR green used showed a 100% selectivity, 100% relative accuracy, 100% relative specificity, 100% relative sensitivity and a limit of detection of 277 and 580 DNA copies for C. botulinum type C and C. botulinum type D, respectively. The method reported here represents a suitable tool for laboratory diagnosis of type C and D botulism and for testing a large number of samples collected during the animal botulism surveillance and prevention activities.     

An analysis of agricultural sustainability of cropping systems in arable and dairy farms in an intensively cultivated plain

In Lombardy region (northern Italy) agricultural systems are mostly based on cereals and forage crops, and are normally intensively cultivated. To monitor and analyse the most important agri-environmental issue of this area (water pollution by nitrates and plant protection products, non-renewable fossil energy exploitation, weed dynamics), we conducted an integrated agronomic, environmental and economic assessment of arable and dairy farming using indicators.The structural and management data about farming and cropping systems were collected by periodic interviews over a 3-year period in seven (three arable and four dairy) representative farms.The nitrogen surplus calculated at field scale ranged from low (27 kg N ha^?1) to high (339 kg N ha^?1) values, depending by the amount of chemical and organic fertilisers applied. Fossil inputs ranged from 11.2 to 46.0 GJ ha^?1; the highest values were due to the high use of machinery and chemical inputs. The efficiency factor in energy transformation (an indicator of the dependence of food and feed production on non-renewable energy) ranged from 5.0 to 12.2. Large variability was also observed for economic performance (gross margin; from -364 to 1078 € ha^?1). The lowest values were observed where the total costs of production (fixed and variable) were elevated. The simplification of cropping systems was responsible for a high probability of weed populations development. The values of Load Index, an indicator describing the potential toxicity of plant protection products on non-target organisms, showed an elevated use of active substances due to the dominance of maize.The indicator-based assessment, founded on a relatively simple data collection procedure, described crop management by combining different aspects into few quantities and highlighted its critical environmental issues. The work represents a starting point for improving cropping system management in the study area; the same procedure could be re-applied if data about improved management scenarios were available.     

Role of the genes Md-ACO1 and Md-ACS1 in ethylene production and shelf life of apple (Malus domestica Borkh)

Shelf life determines the economic life time of mature apples, which can be either freshly harvested or stored. Good shelf life is highly associated with a slow decrease of fruit firmness at room temperature. Apple is a climacteric fruit, in which loss of firmness seems to be physiologically related to ethylene. Ethylenes biosynthetic pathway is controlled by two large gene families coding for 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxydase (ACO).In this study, one ACS and one ACO gene were examined for their effect on ethylene production and shelf life in apple using gene specific molecular marker, and have also been positioned on a molecular marker linkage map. The ACO marker was developed in this research and mapped on linkage group (LG) 10 of the crosses Prima × Fiesta and Fuji × Mondial Gala, within the 5% border of a previously identified fruit firmness QTL [Theor Appl Genet 100 (2000) 1074]. We denoted this locus as Md-ACO1. In addition, we mapped the previously developed Md-ACS1 marker [Theor Appl Genet 101 (2000) 742] on LG15.Studies on the cross Fuji × Braeburn revealed that Md-ACS1 and Md-ACO1 independently affect the internal ethylene concentration (IEC) as well as shelf life of apple, Md-ACS1 having the strongest effect. Descendants homozygous for Md-ACS1-2 and Md-ACO1-1 showed to have the lowest ethylene production as well as superior shelf-life. These two genes are candidates to be included in marker assisted breeding.