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.     

Venom immunotherapy for Hymenoptera allergy in a dog

A 1.5-year-old male castrated dog was presented in anaphylactic shock after suffering an apparent bee sting. Immunotherapy with bee venom was initiated based upon history, skin testing and serological testing for allergen-specific immunoglobulin (Ig)E. The dog was maintained on venom immunotherapy for five years and showed no signs of adverse effects from therapy or from repeated bee stings.     

Pathologic and immunohistochemical findings in an outbreak of systemic toxoplasmosis in a mob of red kangaroos

Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.     

Production,chemical composition,and fatty acid profile of milk from buffaloes fed with cupuaçu (Theobroma grandiflorum) cake and murumuru (Astrocaryum murumuru) cake in the Eastern Amazon

The objective was to evaluate the effect of concentrate supplementation using by-products of the Amazonian industry on milk production, milk composition, and milk fatty acid profile of dairy buffaloes. Twelve lactating buffaloes (544.5 ± 35.6 kg, 6.4 ± 2.2 years old, 59 ± 6 days in milk) were allotted in a pasture of Mombaça grass and managed under rotational grazing (4 days occupancy/28 days rest). A 3 × 3 Latin square was adopted, and each animal alternately received three supplementary treatments based on corn bran + soybean meal or cupuaçu cake or murumuru cake for 21 days per treatment. Murumuru cake increased the levels of lauric acid and myristic acid in the milk (p < 0.05). Murumuru cake reduced the unsaturated fatty acid contents in the milk compared with animals fed control diet or cupuaçu cake (24.27% vs. 25.24% vs. 25.08%). The n-6/n-3 ratio was 2.6, 1.97, and 2.0 in the control, cupuaçu, and murumuru groups, respectively. Based on this parameter, cakes made from cupuaçu as well as murumuru could be considered to be adequate for inclusion in dairy water buffalo feed. However, the murumuru cake addition requires some caution because its use induces the secretion of higher levels of lauric and myristic fatty acids that are related to human cardiovascular disease.     

Describing Human–Wildlife Interaction from a European Perspective

European researchers from both the natural and social sciences show growing interest in studying interactions between society and wildlife. A wealth of theoretical frameworks, concepts, and methods are used, but an integration of perspectives is lacking. This research note summarizes results from two workshops that included 63 delegates from 25 European countries, as well as a follow-up survey of 41 respondents. Two main theoretical approaches to the study of human–wildlife interactions were identified. One approach focuses on the collective societal level relying on theories of governance, social representation, deliberative procedures, and commons theory. The other approach targets individuals or groups, and is based on theories such as the cognitive hierarchy, theory of reasoned action, and theory of planned behavior. Interdisciplinary collaboration is needed to identify the best options for wildlife conservation and management in a more politically integrated Europe.     

Successful management of jejunojejunal anastomosis dehiscence by extra-abdominal exteriorization and bandaging in a cat with septic peritonitis

Duodenal and jejunal resections were performed in a cat with septic peritonitis due to small intestinal perforations by a linear foreign body. Three days later jejunal resection and anastomosis were repeated due to dehiscence of the anastomosis site. This segment of intestine was exteriorized through the body wall and managed with bandages for 5 days before it was surgically replaced into the abdomen. The cat made a full recovery.     

The risk of salmonellae shedding by dogs fed Salmonella-contaminated commercial raw food diets

Twenty-eight research dogs were enrolled to determine the prevalence of salmonellae shedding after consumption of 1 Salmonella-contaminated commercial raw food diet meal. Sixteen dogs were exposed to Salmonella-contaminated commercial raw food diets and 12 to Salmonella-free commercial raw food diets. Seven of the exposed dogs shed salmonellae 1-7 days after consumption of Salmonella-contaminated raw food diets. None of the dogs fed Salmonella-free diets shed salmonellae. No clinical signs were observed in either group. Five of the 7 dogs shed the same serotypes as those recovered from food samples used for feeding. Results showed the same serotypes and antimicrobial resistance pattern in 2 of the 7 shedders. Dogs fed Salmonella-contaminated raw food diets can shed salmonellae and may, therefore, be a source of environmental contamination potentially leading to human or animal illness.     

In Vitro Assessment of the Equine Hoof Wall Strains in Flat Weight Bearing and After Heel Elevation

Horseshoeing is a common practice, but effects on the hoof wall are poorly understood. Strain gauges were used to document and compare hoof behavior in vitro during flat weight bearing and after artificial heel elevation. Ten front limbs of Thoroughbred race horses, shod with conventional flat shoes, were used. Eight strain gauges were symmetrically distributed around the toe, quarters, and heels. Each limb was mounted to a testing machine (Kratos K5002; Kratos Dynamômetros, Ltda., Cotia-SP-Brazil) and subjected to a load equivalent to 30% of the donor's body weight. Strains (μ) were acquired by means of a computerized system and the results compared using Friedman and Wilcoxon statistical tests. There was greater strain variation when the heels were elevated. Compression predominated during flat weight bearing, with a tendency to horizontal traction after heel elevation. The changes in strain caused by heel elevation were not always symmetrical. Elevation of the heels tensed the toe and the medial quarter horizontally, increased load at the posterior portion of the hoof capsule, and hindered its expansion.     

Being exposed to low concentrations of pirimiphos-methyl and chlorfenapyr has detrimental effects on the mobility of Trogoderma granarium

BACKGROUND

Sublethal effects of insecticides may negatively affect several biological and behavioral traits of insects. The lethal effects of pirimiphos-methyl and chlorfenapyr have been previously showed on Trogoderma granarium, but little knowledge is available about their sublethal effects at low concentrations on both sexes. Herein, the sublethal effects of pirimiphos-methyl and chlorfenapyr on the mobility of T. granarium males and females were investigated.

RESULTS

Lethal concentration (LC) values of pirimiphos-methyl and chlorfenapyr were lower for T. granarium females than males. LC values on males were LC₁₀ = 0.000788 and 0.00139 mg active ingredient (a.i.) cm⁻², LC₃₀ = 0.00350 and 0.00535 mg a.i. cm⁻², and LC₅₀ = 0.00986 and 0.0136 mg a.i. cm⁻² for pirimiphos-methyl and chlorfenapyr respectively. LC on females were LC₁₀ = 0.000704 and 0.00110 mg a.i. cm⁻², LC₃₀ = 0.00323 and 0.00428 mg a.i. cm⁻², and LC₅₀ = 0.00925 and 0.0110 mg a.i. cm⁻² for pirimiphos-methyl and chlorfenapyr respectively. The walking duration of beetles exposed to LC₃₀ of pirimiphos-methyl was significantly lower than the individuals exposed to LC₁₀ and LC₃₀ of both insecticides and control ones. Pirimiphos-methyl LC₃₀-exposed males remained more time on their back (101.7 s) than females (46.9 s), while the latter stayed immobile longer than males (381.7 s versus 371.9 s). The highest speed was recorded for control beetles (14.17 mm s⁻¹ females vs. 12.44 mm s⁻¹ males), while the lowest speed was observed in pirimiphos-methyl LC₃₀-treated males (8.36 mm s⁻¹) and females (9.66 mm s⁻¹).

CONCLUSIONS

Overall, males and females exposed to low concentrations of pirimiphos-methyl and chlorfenapyr showed reduced motility. This knowledge can be exploited further to unlock behavioral effects of insecticides for effective pest management programs in warehouses. © 2023 Society of Chemical Industry.