Host specificity and genetic diversity of race 4 strains of Ralstonia solanacearum

Ralstonia solanacearum race 4 isolates were obtained from Zingiberaceae plants in India during bacterial wilt outbreaks. Polyphasic phenotypic and genotypic analysis revealed intraracial diversity and dominance of biovar 3 over biovar 4. Biovar 3 strains were isolated from very severely wilted Zingiberaceae plants in the field and found to be present across diverse geographical, host and seasonal boundaries. It was hypothesized that these isolates belong to a single, ‘fast wilting’, lineage. Using one ‘fast wilting’ isolate in controlled inoculations, rapid wilt was observed in ginger within 5–7 days. Wilting was also observed in several other closely and distantly related hosts such as turmeric (Curcuma longa), aromatic turmeric (Curcuma aromatica), black turmeric (Curcuma caesia), sand ginger (Kaempferia galanga), white turmeric (Curcuma zeodaria), awapuhi (Zingiber zerumbet), greater galangal (Alpinia galanga), globba (Globba sp.), small cardamom (Elettaria cardamomum) and large cardamom (Ammomum subulatum) of the Zingiberaceae family, and in tomato (Solanum lycopersicum). Molecular analysis, including multiplex PCR‐based phylotyping, sequence analysis of 16S rDNA, 16–23S intergenic spacer and the recN gene, and multilocus sequence typing, revealed minimal differences between fast wilting isolates, confirming that almost all belong to the same lineage. Biovar 4 was isolated from plants showing slow wilt progression and self‐limiting wilting in restricted geographical locations instead, and was identified to be genetically distinct from the fast wilting biovar 3 isolates. To the authors' knowledge, this is the first report of host range and genetic analysis of R. solanacearum race 4 in India.     

Association of Pedilanthus leaf curl virus with yellow mottling and leaf curl symptoms in two jasmine species grown in India

Association of Pedilanthus leaf curl virus (PeLCV) with yellow mottling of crape jasmine (Tabernaemontana coronaria) and leaf curl of night blooming jasmine (Cestrum nocturnum) was investigated using a sequence analysis of the cloned full-length genome amplified by Rolling circle amplification using Φ-29 DNA polymerase. In the sequence analysis, virus isolates from T. coronaria (JN807764) and C. nocturnum (JQ012790) shared 95 % sequence identity with each other and the highest identity (93 %) and the closest phylogenetic relationships with PeLCV (AM712436); therefore, the virus isolates were identified as PeLCV strains. This report from India is the first on the occurrence of PeLCV on T. coronaria and C. nocturnum anywhere in the world.     

Isolation, culturing and characterization of feeder-independent amniotic fluid stem cells in buffalo (Bubalus bubalis)

Heterogeneous amniotic fluid contains various cell types. The aim of this study was to characterize and differentiate some of the key stemness attributes of the amniotic fluid-derived cells in buffalo (Bubalus bubalis). The amniotic fluid (AF) cells were cultured without feeder cells, in DMEM containing 15% FBS, 1% non-essential amino acids, 1% penicillin/streptomycin/ampicillin, 1% vitamin solution, and 1% l-glutamine in 5% CO(2) in humidified air at 38.5±0.5 °C. After 6 days of culture different types of cells viz., star shaped (62.7%), spherical without nucleus (1.9%), spherical with nucleus (26.4%), pentagonal (0.4%), and free floating/rounded cells (8.3%) were observed. Most of the cells started anchorage-dependent growth after day 7 of the culture. Expression of alkaline phosphatase (AP) and Oct-4, Nestin and FGF-5 were observed from the AF cells at different passages. Using species-specific primers, a PCR amplicon of 200, 296 and 210 bp were observed for Oct-4, Nestin and FGF-5, respectively. The cells were found to have a normal karyotype at different passages. These results may contribute towards establishing non-embryonic pluripotent stem cells for various therapeutic and reproductive biotechnological applications in the species.     

Nutritional evaluation of organically grown fodders in lactating Murrah buffaloes (Bubalus bubalis)

A study was undertaken to evaluate the nutritional response of organically grown diets in buffaloes on nutrients utilization and nutrient efficiency for milk production. For this, ten milch Murrah buffaloes of average body weight (490.72?±?6.65 kg), milk yield (8.13?±?0.33 kg), and lactation stage (85?±?5.28 days) were distributed in inorganic (InDg₎ and organic dietary groups (OrDg) with five animals in each. Buffaloes of InDg and OgDg were fed sorghum hay–berseem fodder-concentrate mixture-based diets grown inorganically and organically, respectively. After 60 days of feeding, a digestion cum metabolism trial was conducted to assess feed intake, nutrient utilization, and N balance, while milk yield and composition were recorded fortnightly. DM, CP, digestible crude protein (DCP), and metabolizable energy (ME) intake (g/Kg w^0.75) were similar in animals of InDg and OrDg. NDF, ADF, and hemicellulose digestibility were (P?<?0.05) higher in animals on OrDg (59.20, 51.55, and 62.67) than InDg diet (54.57, 43.72, and 56.61 %), respectively. Urinary n loss (g/day) was (P?<?0.05) lower in OrDg (67.23) than in InDg (83.55); however, milk N was comparable in animals of both dietary groups (47.36 vs 45.82 g/day). Nitrogen balance was higher in animals of OrDg (39.72) than in InDg (28.08). DCP, TDN, and ME values of both diets were similar. No effect of diet was observed on milk yield and its composition; however, increased lactation length decreased milk yield. Buffaloes on both diets had similar efficiency and conversion ratio of nutrients for milk production. Results revealed that diets (organically vs. inorganically grown) have no effect on milk yield and its composition; however, buffaloes on organic diet have higher fiber digestibility and low urinary N loss which did not affect the dietary nutrients efficiency and conversion ratio for milk production.     

Impact of norgestomet supplementation during early luteal phase on subsequent luteal profiles and conception rate in buffalo: a preliminary study

The current study was aimed to establish the impact of progesterone supplementation (norgestomet progestagen) between days 4 to 10 post-ovulation on subsequent luteal profile and conception rate in buffaloes. The 28 Murrah buffaloes of second to fourth parity, having normal reproductive organs, were estrus synchronized by double PGF_2α protocol at 11 days apart. The buffaloes were inseminated during mid- to late estrus and thereafter repeated at 24 h interval. The buffaloes were randomly assigned into two groups: (1) control (no treatment, n?=?14) and (2) treatment group (CRESTAR ear implant, n?=?14). The CRESTAR ear implant (3 mg, norgestomet progestagen) was inserted subcutaneous between days 4 to 10 post-ovulation. The ovaries were scanned at estrus and thereafter on days 4, 10, 16, 21, and 40 post-ovulation to examine the preovulatory follicle (POF) and corpus luteum (CL) diameter. Each ultasonography was followed by blood sample collection for analysis of plasma progesterone concentrations following ovulation. The conception rate was similar (p?>?0.05) between treated and control buffaloes. The pregnant buffalo of the control group had larger (p?<?0.05) POF diameter than nonpregnant counterparts. The CL diameter was similar (p?>?0.05) in both treated and untreated control as well as in their pregnant and nonpregnant buffaloes of the respective groups. The plasma progesterone concentrations were higher (p?<?0.05) in the treatment group on the day 10 post-ovulation as compared to the control buffaloes. It is concluded that norgestomet supplementation had no impact on conception rate and CL diameter but enhances the plasma progesterone concentrations following treatment in buffaloes.     

Plasma hormones, metabolites, milk production, and cholesterol levels in Murrah buffaloes fed with Asparagus racemosus in transition and postpartum period

Ten dry and pregnant Murrah buffaloes were selected to investigate the effect of Asparagus racemosus feeding on hormones, metabolites, milk yield, and plasma cholesterol levels. The treatment groups of buffaloes were fed with A. racemosus (shatavari) @?150?g/day/animal during prepartum and @?300?g/day/animal during the postpartum period. Blood samples collected on ?6, ?4, ?2-week, day of parturition (0), and +2, +4, and +6-week postpartum were analyzed for plasma total cholesterol, triglycerides, HDL, low-density lipoproteins (LDL), prolactin, cortisol, and blood metabolites. Milk samples collected at weekly intervals (+1, +3, +5, and 7?weeks) were analyzed for total milk fat cholesterol. Prepartum plasma cholesterol concentrations were significantly higher in treatment group over the control (P?<?0.05). Mean plasma triglycerides, LDL cholesterol, HDL cholesterol, glucose, and nonesterified fatty acid (NEFA) levels varied nonsignificantly between groups. Plasma prolactin and cortisol concentrations were significantly (P?<?0.01) more in treatment group than in control group. On day of parturition, plasma prolactin, cortisol, LDL, and plasma total cholesterol were higher (P?<?0.01) in treatment group buffaloes in comparison to control group. A. racemosus feeding significantly (P?<?0.01) increased plasma prolactin, cortisol (P?<?0.01), and milk fat cholesterol (P?<?0.05) without affecting total cholesterol, HDL, LDL, glucose, and NEFA concentrations. The buffaloes of treatment group produced more milk (?@?0.526?kg/animal/day) suggesting thereby that A. racemosus is galactopoietic. It was concluded that feeding of A. racemosus increases plasma prolactin and cortisol and decreased plasma total cholesterol and LDL concentration.     

Correlations among <Emphasis Type="Italic">Anaplasma marginale</Emphasis> parasitemia and markers of oxidative stress in crossbred calves

The present study was designed to determine the correlations among Anaplasma marginale parasitemia and markers of oxidative stress in crossbred calves. Blood was collected from 11 crossbred calves infected with A. marginale along with 11 healthy crossbred calves as controls for determination of hematology and oxidative stress indicators. Percentage of parasitemia in infected calves varied from 0.8% to 6.0%. The values of hematological indicators and antioxidant enzymes were decreased, whereas erythrocytic lipid peroxidation (LPO) and plasma nitrate (NO) level were significantly (p < 0.05) augmented in A. marginale-infected animals over healthy group. Parasitemia was positively correlated (p < 0.01) with erythrocytic LPO and plasma NO and negatively correlated (p < 0.01) with hematological indicators and antioxidant enzymes. In addition, erythrocytic LPO was negatively correlated (p < 0.01) with the hemoglobin, erythrocyte count, and packed cell volume. From the present study, it can be concluded that anaplasmosis in crossbred calves is associated with a parasitic load-dependent oxidative damage as indicated by poor antioxidant status and enhanced oxidative stress, which are contributed to severe anemia.