Artificial polyploidy in medicinal plants: Advancement in the last two decades and impending prospects

Medicinal plants are in huge demand since the consumption is widespread and ever-increasing globally. The conventional breeding programs are generally environmental dependent; prone to different biotic and abiotic stresses as well as the secondary metabolite content is too low to harvest. In this context, developing polyploid individuals artificially would be a remarkable approach to increase vigor and attain this objective. Polyploids often exhibit some morphological features that are different or greater in forms than their diploid progenies. Polyploidization can be induced by quite a few antimitotic agents. The most frequently used antimitotic chemicals are colchicine, trifluralin, and oryzalin. The whole method of induced chromosome doubling consists of a series of steps, including an induction phase, regrowth phase, and a confirmation technique to evaluate the rate of achievement. The induction phase depends on different factors, such as explant types, antimitotic agents, its different concentrations, and exposure durations. To evaluate the accomplishment of polyploidization, morphological or anatomical observations are recorded as a rapid method. However, chromosome count and flow cytometry are the most eminent method for absolute confirmation. Despite significant prospects of polyploidization, there has been very little research on medicinal plants. The current review gives an overview of the different parameters of in vitro chromosome doubling, the history of the technique, and progress made over the last two decades.     

Effect of supplementation of mustard oil cake on intake,digestibility and microbial protein synthesis of cattle in a straw-based diet in Bangladesh

The objective of this study was to analyse the effects of different levels of rumen-degradable protein (RDP) on intake, digestibility and microbial protein synthesis by supplementing mustard oil cake (MOC) on rice straw-based diet of cattle (Bos indicus) in Bangladesh. A 4 × 4 Latin square design was applied. Four diets having constant energy (7.0 MJ/kg of dry matter (DM)) with varying levels of RDP (M ₀ = 4.1 g/MJ (control), M ₁ = 6.3 g/MJ, M ₂ = 8.3 g/MJ and M ₃ = 12.4 g/MJ of metabolizable energy (ME)) were received by each animal for a period of 28 days. A metabolism trial was conducted for 7 days. Results indicate that with increasing levels of RDP, crude protein (CP) and RDP intake increased significantly (P < 0.01). The significant (P < 0.01) increase in digestibility values are obtained for DM, organic matter, CP and digestible organic matter in the rumen. The digestibility of neutral detergent fibre and acid detergent fibre was also increased significantly (P < 0.05). The total nitrogen (N), ammonia-N and total volatile fatty acids increase significantly (P < 0.01) while the rumen pH increased from M ₀ to M ₂ and decreased thereafter. The efficiency microbial N intake increased significantly (P < 0.01) but showed a curvilinear response with higher RDP level (12.40 g/RDP/MJ ME). This study concludes that supplementation of RDP from MOC enhances the intake, digestibility and microbial protein synthesis which ultimately increases utilization of low-quality feed resources that can be used for developing cost-effective feeding systems on a straw-based diet in tropical regions.     

The changes in physiological and biochemical traits of Tibetan wild and cultivated barley in response to low phosphorus stress

Low phosphorus (LP) limits crop growth and productivity in the majority of arable lands worldwide. Here, we investigated the changes in physiological and biochemical traits of Tibetan wild barleys (Hordeum vulgare L. ssp. spontaneum) XZ99 (LP tolerant), XZ100 (LP sensitive), and cultivated barley ZD9 (moderately LP tolerant) under two phosphorus (P) levels during vegetative stage. These genotypes showed considerable differences in the change of biomass accumulation, root/shoot dry weight ratio, root morphology, organic acid secretion, carbohydrate metabolism, ATPase (Adenosine triphosphatase) activity, P concentration and accumulation under LP in comparison with CK (control) condition. The higher LP tolerance of XZ99 is associated with more developed roots, enhanced sucrose biosynthesis and hydrolysis of carbohydrate metabolism pathway, higher APase (Acid phosphatase) and ATPase activity, and more secretion of citrate and succinate in roots when plants are exposed to LP stress. The results prove the potential of Tibetan wild barley in developing barley cultivars with high tolerance to LP stress and understanding the mechanisms of LP tolerance in plants.     

Comparison of direct immunofluorescence, immunoassays, and fecal flotation for detection of Cryptosporidium spp. and Giardia spp. in naturally exposed cats in 4 Northern California animal shelters

BACKGROUND: Giardia spp. and Cryptosporidium spp. are common intestinal protozoan parasites in domestic cats. Few studies have critically evaluated the performance characteristics of commercially available immunoassays for detection of these organisms in the cat. HYPOTHESIS: Human-based immunoassays are suboptimal for the detection of Giardia spp. and Cryptosporidium spp. in cats. ANIMALS: Three-hundred-and-forty-four cats with diarrheic and nondiarrheic fecal specimens at 4 northern California animal shelters. METHODS: A fecal specimen was collected from each cat in a case-controlled fashion. Fecal specimens were tested for Giardia spp. and Cryptosporidium spp. by using centrifugation flotation and 5 commercially available immunoassays (SNAP Giardia, ProSpecT Giardia Microplate Assay, ProSpecT Cryptosporidium Microplate Assay, ImmunoCard STAT! Cryptosporidium/ Giardia Rapid Assay, and Xpect Giardia/Cryptosporidium). Results were compared with a reference standard, the MeriFluor direct immunofluorescence assay. RESULTS: Overall prevalences of Giardia spp. and Cryptosporidium spp. were 9.8 and 4.7%, respectively. The ProSpecT Microplate Assay had the highest sensitivities and specificities for Giardia spp. (91.2 and 99.4%) and Cryptosporidum spp. (71.4 and 96.7%), respectively. The SNAP Giardia antigen assay was easier to use and equally sensitive (85.3%) and specific (100%) to fecal flotation. CONCLUSIONS AND CLINICAL IMPORTANCE: Caution should be exercised when using human-based immunoassays for the diagnosis of Giardia and Cryptosporidium spp. in cats. Fecal flotation remains a useful method for detection of Giardia spp., can be used to detect other parasites, and has a sensitivity of 97.8% for detection of Giardia spp. when combined with the SNAP Giardia immunoassay.     

Zoonotic pathogens isolated from wild animals and environmental samples at two California wildlife hospitals

During 2010, 48 states and Puerto Rico reported 6,154 rabid animals and 2 human rabies cases to the CDC, representing an 8% decrease from the 6,690 rabid animals and 4 human cases reported in 2009. Hawaii and Mississippi did not report any laboratory-confirmed rabid animals during 2010. Approximately 92% of reported rabid animals were wildlife. Relative contributions by the major animal groups were as follows: 2,246 raccoons (36.5%), 1,448 skunks (23.5%), 1,430 bats (23.2%), 429 foxes (6.9%), 303 cats (4.9%), 71 cattle (1.1 %), and 69 dogs (1.1 %). Compared with 2009, number of reported rabid animals decreased across all animal types with the exception of a 1 % increase in the number of reported rabid cats. Two cases of rabies involving humans were reported from Louisiana and Wisconsin in 2010. Louisiana reported an imported human rabies case involving a 19-year-old male migrant farm worker who had a history of a vampire bat (Desmodus rotundus) bite received while in Mexico. This represents the first human rabies case reported in the United States confirmed to have been caused by a vampire bat rabies virus variant. Wisconsin reported a human rabies case involving a 70-year-old male that was confirmed to have been caused by a rabies virus variant associated with tri-colored bats (Perimyotis subflavus).     

Evaluation of five enzyme immunoassays compared with the cytotoxicity assay for diagnosis of Clostridium difficile-associated diarrhea in dogs.

Clostridium difficile-associated-diarrhea (CDAD) is a nosocomial infection in dogs. Diagnosis of this infection is dependent on clinical signs of disease supported by laboratory detection of C. difficile toxins A or B, or both, in fecal specimens via enzyme-linked immunosorbent assay (ELISA). Unfortunately, to the authors' knowledge, commercially available ELISAs have not been validated in dogs to date. We evaluated 5 ELISAs done on 143 canine fecal specimens (100 diarrheic and 43 nondiarrheic dogs) and on 29 C. difficile isolates. The results of each ELISA were compared with the cytotoxin B tissue culture assay (CTA). Clostridium difficile was isolated from 23% of the fecal specimens. Eighteen of the 143 fecal specimens were toxin positive (15 diarrheic and 3 nondiarrheic dogs). On the basis of multiplex polymerase chain reaction (PCR) analysis for toxin-A and -B genes, 72% of the isolates were toxigenic. The carriage rate of toxigenic isolates in diarrheic dogs was higher than that in the nondiarrheic dogs; however, these differences were not statistically significant. A good correlation was found between CTA, PCR, and culture results. The ELISAs done on fecal specimens collected from diarrheic dogs had low sensitivity (7-33%). In contrast, ELISA for toxin A or B, or both, performed on toxigenic isolates had high sensitivity (93%). These results suggest that commercially available human ELISAs are inadequate for the diagnosis of canine C. difficile-associated diarrhea when tested on fecal specimens. In contrast, the Premier ToxinA/B and Techlab ToxinA/B ELISAs may be useful for the diagnosis of canine CDAD when used on toxigenic isolates.