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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 (< 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, < 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.  相似文献   
3.
Blood transfusions in veterinary medicine have become increasingly more common and are now an integral part of lifesaving and advanced treatment in small and large animals. Important risks associated with transfusion of blood products include the transmission of various infectious diseases. Several guidelines suggest what infectious agents to screen for in canine and feline transfusion medicine. However, while the risk of bacterial contamination of blood products during storage and administration has not been documented in veterinary medicine, it has emerged as a cause of morbidity and mortality in human transfusion medicine. Clinical experience shows that the majority of blood component bacterial contaminations are caused by only a few species. Unlike other types of bacteria, psychrotolerant species like Pseudomonas spp. and Serratia spp. can proliferate during the storage of blood units at 4°C from a very low titer at the time of blood collection to a clinically significant level (> 105 CFU/mL) causing clinical sepsis resulting from red blood cell concentrate transfusions in human medicine. The purpose of this report was to describe the detection and quantification procedures applied in 4 cases of bacterial contamination of canine and feline blood units, which suggest the need for further investigations to optimize patients’ safety in veterinary transfusion medicine.  相似文献   
4.

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.  相似文献   
5.

Background

Although the pupal parasitoid Trichopria drosophilae is used in conservative and augmentative biocontrol of Drosophila suzukii infestations, current pest management strategies mostly rely on multiple insecticide applications. In this context, the aim of the study was to investigate the baseline toxicity of nine insecticides on D. suzukii larvae and their multiple sublethal effects (LC10) on immature stages of the pest feeding on contaminated diet and T. drosophilae developing within the intoxicated host.

Results

Chlorpyriphos and azadirachtin showed the lowest and the highest LC10, the values of which were 9.78 × 1013 and 1.46 × 103 times lower than their recommended label field rate, respectively. Among tested insecticides, imidacloprid, malathion and dimethoate were the only treatments that did not affect the juvenile development time of D. suzukii, while spinosad and the organophosphates chlorpyriphos and dimethoate did not influence fly pupal size. No sublethal effects were recorded on T. drosophilae degree of infestation (DI) and juvenile development time. On the contrary, cyazypyr and dimethoate negatively affected the success of parasitism (SP) and the number of progeny of the pupal parasitoid, in association with malathion for the first parameter and spinosad for the fertility. Compared to the untreated control, more female progeny emerged following azadirachtin exposure, while dimethoate caused the opposite effect. Imidacloprid, lambda-cyhalothrin and spinetoram decreased hind tibia length of emerged parasitoids.

Conclusion

This study provides new insights on the (eco)toxicological profile of nine insecticides and new information needed to support the deployment of T. drosophilae in the field within the sustainable management techniques against D. suzukii. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.  相似文献   
6.
Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.  相似文献   
7.
Biofilm-forming ability is increasingly being recognized as an important virulence factor in several Staphylococcus species. This study evaluated the biofilm-forming ability of sixty canine derived clinical isolates of S. pseudintermedius, using three phenotypic methods, microtiter plate test (MtP), Congo red agar method (CRA) and tube adherence test, and the presence and impact of biofilm-associated genes (icaA and icaD). The results showed that icaA and icaD genes were detected concomitantly in 55 (91.7%) of 60 isolates. A majority (88.3%) of the strains screened had matching results by the tube adherence test, MtP and PCR analysis. Better agreement (95%) was found between the PCR-based analysis and the CRA. Results of the icaA and icaD gene PCRs showed good agreement with CRA results, with a kappa of 0.7. Comparing the phenotypic methods, the statistical analysis showed that the agreement among the phenotypical tests using categorical data was generally good. Considering two classes (biofilm producer and biofilm non-producer), the percentage of matching results between the CRA method and the tube adherence test and between the CRA method and the MtP was 93.3%. A concordance of 100% was revealed between the MtP and the tube adherence test. The results indicate a high prevalence of the ica genes within S. pseudintermedius isolates, and their presence is associated with in vitro formation of a biofilm. A combination of phenotypic and genotypic tests is recommended for investigating biofilm formation in S. pseudintermedius.  相似文献   
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alpha(1)-Acid glycoprotein modulates apoptosis in bovine monocytes   总被引:1,自引:0,他引:1  
alpha(1)-Acid glycoprotein (AGP, orosomucoid) is a normal constituent of bovine blood. AGP is an immunocalin, a binding protein that can also exert several immunomodulatory functions. In this paper we investigated the effect of bovine alpha(1)-acid glycoprotein (boAGP) on spontaneous and staurosporine-induced apoptosis of blood derived monocytes purified using magnetic cell sorting techniques. Bovine AGP was purified from blood following a chromatographic protocol. The homogeneous protein was used to stimulate the cells as well to raise a polyclonal antibody, that was used throughout all the experiments. When monocytes were incubated with high concentrations of boAGP (0.9 mg/ml), similar to those found in bovine plasma during systemic reaction to inflammation, their spontaneous apoptosis rate was suppressed, as determined by caspase-3/7 enzymatic activity assay. Similar results were obtained when apoptosis was induced by adding staurosporine, a potent protein kinase inhibitor. The apoptosis-modulating activity of boAGP was dependent on its concentration, since physiological concentrations of boAGP (0.3 mg/ml) did not exhibit a statistically significative anti-apoptotic activity. We also investigated whether this apoptosis-modulating activity was dependent on the terminal sialic acid residues exposed on the surface of the protein. Enzymatic treatment with neuraminidase, that cleaves terminal sialic acid residues, completely abolished boAGP's anti-apoptotic activity. These results suggest that the protective effect of AGP is likely mediated by its sialic acid terminal groups.  相似文献   
10.
Major sugar and carboxylic acid components in apricot flesh fruits were detected and quantified. Fifty-one genotypes including clones growing in France, Spain, Italy, Greece and USA, belonging to the INRA germ plasm collection has been evaluated. Principal component analysis (PCA) has been performed to study correlation among fruit quality measurements and to interpret relationships between genotypes as a tool for germ plasm characterization. A marked variation in malic and citric acid content has been observed and PCA revealed clusters of apricot genotypes for the malic/citric ratio content. A good correlation between sugar component and refractometer index (r = 0.83) has been detected. The wide range of diversity in malic and citric acid content in apricot germ plasm makes it possible to breed and select genotypes with improved flavour on the basis of superior phenotypes. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   
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