Towards developing resistance to chickpea chlorotic dwarf virus through CRISPR/Cas9-mediated gene editing using multiplexed gRNAs

Journal of Plant Diseases and Protection - Mastreviruses are an emerging group of viruses transmitted by leafhoppers and infect both monocot and dicot plants. Chickpea chlorotic dwarf virus...     

Origin of Clostridium perfringens isolates determines the ability to induce necrotic enteritis in broilers

Since the ban on growth-promoting antibiotics in animal feed in the European Union, necrotic enteritis has become a major cause of mortality in broiler chickens. Despite the importance of the disease, the pathogenesis is still not completely understood. In the current study, Clostridium perfringens strains isolated from healthy flocks and isolates from outbreaks of necrotic enteritis were evaluated for the ability to cause gut necrosis in an intestinal loop model in laying hens and in an experimental infection model in broilers. High, intermediate and low alpha toxin producing strains were chosen from each isolation source. Only the isolates from field outbreaks induced necrotic gut lesions, independent of the amount of alpha toxin produced in vitro. It was also shown that alpha toxin producing isolates from calf hemorrhagic enteritis cases were not able to induce necrotic enteritis in poultry. These results suggest the presence of host specific virulence factors in C. perfringens strains, isolated from chickens with intestinal necrotic enteritis lesions.     

Pharmacokinetics of Enrofloxacin and Its Metabolite Ciprofloxacin after Intramuscular Administration of Enrofloxacin in Goats

The pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin were investigated in goats after a single intramuscular administration of enrofloxacin at 2.5 mg/kg body weight. The plasma concentrations of enrofloxacin and ciprofloxacin were determined simultaneously by a HPLC method. The peak concentrations (C _max) of enrofloxacin (1.13 g/ml) and ciprofloxacin (0.24 g/ml) were observed at 0.8 and 1.2 h, respectively. The elimination half-life (t _1/2), volume of distribution (V _d(area)), total body clearance (Cl_B) and mean residence time (MRT) of enrofloxacin were 0.74 h, 1.42 L/kg, 1329 ml/h per kg and 1.54 h, respectively. The t _1/2, area under the plasma concentration–time curve (AUC) and the MRT of ciprofloxacin were 1.38 h, 0.74 g h/ml and 2.73 h, respectively. The metabolic conversion of enrofloxacin to ciprofloxacin was appreciable (36%) and the sum of the plasma concentrations of enrofloxacin and ciprofloxacin was maintained at or above 0.1 g/ml for up to 4 h. Enrofloxacin appears to be useful for the treatment of goat diseases associated with pathogens sensitive to this drug.     

The Blonde d’Aquitaine T3811>G3811 mutation in the myostatin gene: association with growth,carcass, and muscle phenotypes in veal calves

The mutation T3811 → G3811 (TG3811) discovered in the myostatin gene of the Blonde d’Aquitaine breed is suspected of contributing to the outstanding muscularity of this breed. An experiment was designed to estimate the effect of this mutation in an F2 and back-cross Blonde d’Aquitaine × Holstein population. By genotyping all known mutations in the myostatin gene, it was ensured that the TG3811 mutation was indeed the only known mutation segregating in this population. Fifty-six calves (43 F2, 13 back-cross) were intensively fattened and slaughtered at 24.0 ± 1.4 wk of age. The effects of the mutation were estimated by comparing the calves with the [T/T] (n = 18), [T/G] (n = 30), and [G/G] (n = 8) genotypes. Highly significant substitution effects (P < 0.001), above + 1.2 phenotypic SD, were shown on carcass yield and muscularity scores. Birth weight (P < 0.001) was positively affected by the mutation (+0.8 SD) but not growth rate (P = 0.97), while carcass length (P = 0.03), and fatness (P ≤ 0.03) were negatively affected (–0.5 to –0.7 SD). The characteristics of the Triceps brachii muscle were affected by the mutation (P < 0.001), with lower ICDH activity (oxidative) and a higher proportion of myosin type 2X muscle fibers (fast twitch). The effects of the TG3811 mutation were similar to those of other known myostatin mutations, although the Blonde d’Aquitaine animals, which are predominantly [G/G] homozygous, do not exhibit extreme double muscling.     

On the Current Situation of Glanders in Various Districts of the Pakistani Punjab

Glanders is a highly infectious and zoonotic disease of solipeds caused by Burkholderia mallei. Progressive loss of efficiency and fatal outcome resulted in massive economic losses, which forced veterinary authorities throughout the world to implement disease control measures; these measures included mass testing using the complement fixation test and/or malleinization, and the culling of positives. This led to the eradication of glanders from Western Europe and North America in the 1950s. However, in the last decade, the number of outbreaks in Asia and South America increased steadily, and glanders regained the status of a re-emerging transboundary disease. Pakistan has been an endemic country for the past 120 years, but concise data on the presence of disease are not available. A total of 533 serum samples were collected from draught equines, a suspected risk group for glanders, from various districts of Punjab in Pakistan. The complement fixation test and the highly sensitive Western blot technique were used for serodiagnosis. No animal (horse, mule, and donkey) was found to be positive for infection. Glanders seems to be restricted to remote, sporadic pockets of endemicity and may cause outbreaks after being introduced into naive populations by (asymptomatic) shedders.     

Ability of induced corpora lutea to maintain pregnancy in beef cows

Experiments were conducted in beef cows without a primary CL, in which pregnancy had been maintained with exogenous progestogen. In preliminary trials, replacement CL induced ipsilateral to the embryo and after, rather than before, d 36 of pregnancy, maintained more pregnancies after withdrawal of exogenous progestogen (13/13 vs 2/6; P < 0.05). In Exp. 1, in cows with replacement CL induced by treatment with hCG on d 28 of pregnancy, treatment with flunixin meglumine on d 31 through 37 did not increase maintenance of pregnancy. Experiment 2 was conducted to evaluate directly the effects of concentrations of PGF2alpha and estradiol-17beta during d 31 through 35 of pregnancy on maintenance of pregnancy by replacement CL induced between d 28 and 31. In cows that maintained pregnancy while progestogen was provided, maintenance of pregnancy after withdrawal of exogenous progestogen tended to be greater with high (5/5) than with low (2/6; P < 0.10) concentrations of PGF2alpha and greater with low (6/7) than with high (2/6; P = 0.10) concentrations of estradiol-17beta. Secretion of progesterone by replacement CL was greater (P < 0.05) in cows with high than in those with low concentrations of PGF2, during d 31 through 35. Prostaglandin F2alpha may facilitate attachment of the bovine embryo (d 30 to 40) in a manner similar to that reported for implantation in other species. Cows that did not form CL in response to hCG on d 28 to 31 responded well when retreated after d 36. Again, maintenance of pregnancy was greater when replacement CL were induced after (9/9) rather than before d 36 (8/16; P < 0.05).     

Isolation and partial characterization of excretory/secretory antigens of Gastrothylax crumenifer

The present study was carried out to identify the excretory/secretory (E/S) antigens of the rumen infecting digenetic trematode Gastrothylax crumenifer that may be useful for the immunodiagnosis of rumen amphistomosis particularly during the pre-monsoon season during which this rumen parasite stops shedding eggs. The in vitro released E/S proteins were purified on a Sephadex G-200 column. The gel filtration profile revealed three distinct fractions F1-F3 where F1 and F3 appeared as sharp peaks while the F2 fraction was dispersed. The antibody titre against each of the purified E/S fractions was determined by ELISA using anti-whole E/S polyclonal antibodies raised in rabbit. Among the three fractions, the antibody titre against F1 was highest (1:12,800) whereas IgG titre was very low (1:50) for fraction F2 and F3 (1:100). Of the total polypeptides resolved on gradient SDS-PAGE, only a few antigenic polypeptides were detected in each fraction with hyperimmune anti-serum as revealed by Western Blot analysis. However, a 33 kDa antigen detected in each fraction appeared to be immunodominant which could be exploited for the diagnosis of the pouched amphistome.     

Effect of root applied 28-homobrassinolide on the performance of Lycopersicon esculentum

The roots of 20 days old seedlings of tomato (Lycopersicon esculentum Mill.) at the time of their transplantation, were dipped in 10⁻⁸, 10⁻⁷, 10⁻⁶ or 10⁻⁵ M of 28-homobrassinolide (HBR) for 15, 30 or 45 min and were allowed to grow in earthen pots, in a net house. The leaves of the plants, at days 30 and 60, possessed elevated quantities of nitrate reductase (NR), carbonic anhydrase (CA) and the contents of chlorophyll. The values for all the above characteristics were significantly higher than that of the water-fed control. The fruits borned at the treated plants were more in number and possessed a lower quantity of ascorbic acid than the control. Moreover, the fruits at ripening, had higher levels of lycopene and β-carotene. Among the treatments, 15 min feeding of 10⁻⁸ M HBR proved best.     

Pyrrolizidine alkaloids from Senecio lorenthii.

The isolation and NMR spectra of otosenine (1) and seneciphylline (2) from Senecio lorenthii are reported.