Evidence of novel Treponema phylotypes implicated in contagious ovine digital dermatitis and association of treponemes with major lameness causing foot pathogens.

In this study 269 swabs collected from 254 ovine foot lesions and 15 apparently healthy ovine feet were screened by PCR for the presence of major lameness causing foot pathogens viz. Treponema species, D. nodosus, F. necrophorum and T. pyogenes with the presumption that ovine foot lesion positive for Treponema species alone or in association with other three pathogens were categorized as contagious ovine digital dermatitis (CODD). While samples positive for D. nodosus alone or its combination with F. necrophorum and T. pyogenes were considered as footrot (FR) and samples in which F. necrophorum or T. pyogenes was found either alone or in combination were considered as interdigital dermatitis (ID). The overall occurrence of Treponema sp. in ovine foot lesions was 48.0%, and ranged from 33 to 58%. In Treponema positive samples D. nodosus, F. necrophorum and T. pyogenes were present in 34 (27.4%), 66 (54.4%) and 84 (68.5%) in contrast to Treponema negative samples in which these were present in 15 (11.1%), 20 (14.12%) and 17 (12.6%) samples, respectively. The data signifies that Treponema sp. are significantly associated with these foot pathogens and their different combinations with Treponema sp. influence the severity of CODD lesion. The identification of Treponema phylotypes was done by sequencing the 16S rRNA gene fragment of ten representative samples. Out of ten sequences, four (Trep-2, Trep-4, Trep-7 and Trep-10) were identical to Treponema sp. phylotype 1 (PT1) that belongs to phylogroup T. refringens-like, one sequence (Trep-1) was genetically close (90% sequence homology) to Treponema brennaborense while five sequences (Trep-3, Trep-5, Trep-6, Trep-8 and Trep-9) matched with uncultured bacterium clones of treponemes forming separate monophyletic group in phylogenetic tree and could represent new digital dermatitis phylogroup presently containing five ovine specific phylotypes. This is the first report on the presence of Treponema phylotypes other than three digital dermatitis (DD) Treponema phylogroups viz. T. phagedenis-like, T. medium/T. vincentii-like, and T. pedis-like that are frequently detected in CODD lesions. Metagenomic analysis of two representative samples revealed the abundance of genus Treponema in CODD lesion while this genus was absent in swab collected from clinically healthy foot suggesting that it might play primary role in producing CODD. These findings may further aid in understanding the etiopathogenesis of CODD and could help to develop appropriate treatment and mitigation strategies to combat the disease.

Biochemical and proteomic insights revealed selenium priming induced phosphorus stress tolerance in common bean (Phaseolus vulgaris L.).

BACKGROUND: Mineral stress is one of the dominating abiotic stresses, which leads to decrease in crop production. Selenium (Se) seed priming is a recent approach to mitigate the plant's mineral deficiency stress. Although not an essential element, Se has beneficial effects on the plants in terms of growth, quality, yield and plant defense system thus, enhancing plant tolerance to mineral deficiency. METHODS AND RESULTS: The present research was accomplished to find out the effect of Se priming on common bean plant (SFB-1 variety) under phosphorus (P) stress. The seeds were grown invitro on four different MGRL media which are normal MGRL media as control with non-Se primed seeds (Se- P+), non -Se primed seeds grown on P deficient MGRL media (Se- P-), Se primed seeds grown on normal MGRL media (Se+P+) and Se primed seeds grown on P deficient MGRL media (Se+P -). The various morphological and biochemical parameters such as proline content, total sugar content, polyphenols and expression of proteins were analyzed under P stress. The results showed that Se priming has significantly (p ≤ 0.05) affected the morphological as well as biochemical parameters under normal and P stress conditions. The morphological parameters-length, weight, number of nodes and leaves of Se+P+, Se+P- root and shoot tissue showed significant increase as compared to Se-P+, Se-P-. Similarly various biochemical parameters such as total chlorophyll content, proline, total sugar content and polyphenols of Se+P+, Se+P- increased significantly as compared to Se-P+, Se-P-. The differential protein expression in both Se+P+, Se+P- and Se-P+, Se-P- plants were determined using MALDI-MS/MS. The differentially expressed proteins in Se+P+, Se+P- plants were identified as caffeic acid-3-O-methyltransferase (COMT) and SecA protein (a subunit of Protein Translocan transporter), and are found responsible for lignin synthesis in root cell walls and ATP dependent movement of thylakoid proteins across the membranes in shoot respectively. The differential expression of proteins in plant tissues, validated morphological and biochemical responses such as maintaining membrane integrity, enhanced modifications in cellular metabolism, improved polyphenol activities and expression of defensive proteins against mineral deficiency. CONCLUSIONS: The study provided an understanding of Se application as a potential approach increasing tolerance and yield in crop plants against mineral deficiency.

Removal of an agricultural herbicide (2,4-Dichlorophenoxyacetic acid) using magnetic nanocomposite: A combined experimental and modeling studies.

This study focused on modeling the removal of one of the widely used agricultural herbicides known as 2,4-Dichlorophenoxyacetic acid (2,4-D) using polypyrrole-coated Fe2O3 nanoparticles (Fe2O3@PPy). The Fe2O3@PPy nanocomposite was synthesized by surface-coating the Tabebuia aurea leaf extract synthesized Fe2O3 nanoparticles with polypyrrole. After characterization, the adsorptive potential of the nanocomposite for removing 2,4-D from aqueous solution was examined. Central composite design (CCD) was employed for optimizing the adsorption, revealing an adsorption efficiency of 90.65% at a 2,4-D concentration of 12 ppm, a dosage of 3.8 g/L, an agitation speed of 150 rpm, and 196 min. Adsorption dataset fitted satisfactorily to Langmuir isotherm (R2: 0.984 & χ2: 0.054) and pseudo-second-order kinetics (R2: 0.929 & χ2: 0.013) whereas the exothermic and spontaneous nature were confirmed via the thermodynamic study. The predictive models, including adaptive neuro-fuzzy inference system (ANFIS), artificial neural network (ANN), and response surface methodology (RSM), demonstrated good precision for the prediction of 2,4-D adsorption, with respective R2 of 0.9719, 0.9604, and 0.9528. Nevertheless, statistical analysis supported ANFIS as the better forecasting tool, while RSM was the least effective. The maximum adsorption capacity of 2,4-D onto the Fe2O3@PPy nanocomposite was 7.29 mg/g, significantly higher than a few reported values. Therefore, the Fe2O3@PPy nanocomposite could serve as a competent adsorbent to remove 2,4-D herbicide from aqueous streams.

Fusarium flocciferum, causing wilt disease of chili (Capsicum annuum L.) in Northern Himalayas: A first report.

In August 2020 chili (Capsicum annuum L.) showing wilt symptoms were collected from different districts of the Kashmir: Pulwama, Srinagar, Baramulla, and Anantnag. From each district one location was selected for sample collection and a total of 23 chili isolates were isolated. The tissue bit technique was used to isolate fungus from the infected samples on potato dextrose agar (PDA) medium, purified using the single spore technique, maintained at 25°±1℃ and then stored at 4° C (Ferniah et al. 2014) . Initially cultural characteristics appeared as white colonies which gradually turned to pale white colored and attained a growth of 90 mm in 18 days of incubation at 25 ± 1°C. Microscopic observations revealed that mycelium was branched and cylindrical, 3.53-4.98 µm in width. Microconidia were ellipsoidal, hyaline, 0-1 septa werepresent, and 6-7 x 3-4 µm in size. Macroconidia were cylindrical, hyaline, 2-6 septa, measuring 20-60 x 40-45 µm in size. Molecular identification of the pathogens with ITS, TEF, and RPB2 was successfully carried out and the fungi was confirmed as Fusarium flocciferum infecting chili. Amplified PCR products were sequenced and were successfully submitted and accessioned in GenBank with accession number OM189458, OM441199, OR484037 for ITS, TEF, and RPB2 gene. To confirm Koch's postulates pathogenicity test was carried out using rhizosphere inoculation technique (Najar et al. 2011, Parihar et al. 2022). In total 7 replications for sand maize meal medium (potting mixture) was prepared by autoclaving 90 g of sand and 10 g of maize meal in 250 ml of erlenmeyer flask comprising 40 ml of distilled water. The spore suspension at 100 µl per pot was inoculated and was mixed with the sterilized potting mixture in a ratio of (2:1) and up to seven days pathogen was allowed to infect the soil (Davey and Papavizas 1962; Hami et al. 2021). Then chili seeds (cv. Kashmir long-1) were sown in infected potting mixture and grown for three weeks to allow the pathogen to infect the host plants. F. flocciferum took six weeks for appearance of symptoms in the infected potted plants. Control mock inoculation of the potting mixture was carried out using water droplets instead of spore suspension at 100 µl per pot. Seven replications were kept for both inoculated and un-inoculated / control mock pots. The plants developed initial symptoms from light green to yellowish discoloration of leaves followed by the drooping, shriveling, and ultimately leading to death. The collar region of the plant was cut vertically and observed that vascular bundles showed brownish spots and discoloration, indicating wilt as the cause of death. The pathogens were re-isolated and inoculated from all infected plants, then compared with their original pure culture inoculated first, which completely resembled based on morphological, cultural, and pathogenic characteristics. No symptoms were observed on control plants. A phylogenetic analysis was also carried out using ClustalW software that grouped the species identified by different genes into different clades. F. flocciferum has been reported earlier in pea, faba bean and bamboo (Kainthola et al. 2022; Sisic et al. 2020) . In solanaceous crops, this species have been explored as wilt pathogens for the first time from India, indicating diversifying nature of Fusarium flocciferum across various hosts including solanaceous crops.

An inexpensive phytoremediation system for treating 50,000 L/day of sewage.

The paper describes the setting up and long-term continuous operation of the first real-life, pilot scale, sewage treatment plant based on the recently patented phytoremediation technology, trademarked as SHEFROL®. The unit was about three times cheaper to install, operate and maintain than the least expensive of the other wetland-based technologies presently in vogue. Its semi-permanent version is 30 times cheaper. Monitoring of flow rates and levels of treatment intermittently over a 3 year course of continuous operation indicated the constancy and robustness of the reactor in treating total solids, suspended solids, chemical oxygen demand, biological oxygen demand, total Kjeldahl nitrogen, and soluble phosphorous to the average extents of 94, 84, 79, 70, 62 and 28% respectively. Earlier experience with bench-scale SHEFROL® units has indicated that removal of metals like Cu, Ni, Co, Zn, and Mn also takes place to the extent of 25-45% in these systems. These primary, secondary, and tertiary treatments occurred in a single unit process with no necessity of any pumping, aeration, or recycling. Models based on artificial intelligence were developed which enable forecasting the reactor performance in terms of secondary and tertiary treatment, respectively.

The paper describes the setting-up, long-term (3 years) performance evaluation at pilot scale, and modeling of a recently patented novel and unprecedented phytoremediation-based sewage treatment technology. The system is seen to be much more robust, versatile, space efficient, and economical, than existing macrophyte-based sewage treatment systems.

Earth observation satellite data-based assessment of wetland dynamics in the Kashmir Himalaya.

Earth observation (EO) technology offers enormous opportunities to assess the magnitude and patterns of spatial variability in wetlands over time. This study aims to assess the spatial and temporal changes in the wetlands of the Kashmir valley using multiple remote sensing satellite data products, Geographic Information System (GIS), and field observations. Moreover, role of major factors operating at different time scales including regional geology, climate, and human activities in driving the wetland change is presented. The dynamics of the wetlands are illustrated in the occurrence, seasonality, and recurrence of surface water, land cover transitions and loss patterns particularly for the period from 1984 to 2021. Constituting about 3% (495 Km2) of the total area, substantial and variable patterns of seasonal and annual changes are exhibited by the wetlands. The main transitions of the water surface reveal that 2% of the area has changed from permanent to seasonal; 8% is lost; 15% is new seasonal; 0.12% is permanently lost; and 0.3% is new permanent. About 22% of the area reveals increase in the intensity of water surface occurrence, whereas 44% shows no change, and 34% exhibits decrease. Bathymetric analysis suggests that the average depth of the wetlands ranges between 0.6 and 16.6 m. In general, alpine wetlands are relatively deeper and mostly static in their structure whereas those in the floodplain are shallow, fragmented, and showing signs of depletion during the assessment period. The results of this assessment will inform the policy on conservation and sustainability of wetlands in the Kashmir Himalaya.

DNA marker based diversity across rice genotypes and advanced breeding lines bred for temperate regions of North-West India.

BACKGROUND: Characterization and evaluation of plant genetic resources play an important role for their utilization in the crop improvement programmes. METHODS AND RESULTS: This study involves the agro-morphological and cooking quality besides, molecular characterization of 51 genotypes/advance breeding lines of rice from Kashmir Himalayas. Significant variability was observed for all agro-morphological and cooking quality traits among all the studied genotypes. Cluster analysis using UPGMA method divided the genotypes into two major clusters having 15 and 36 genotypes. Thirty eight genotypes screened using 24 SSR markers detected 48 alleles with 2.0 alleles for each locus with average polymorphism information content (PIC) of 0.37. High polymorphism information content (PIC) values was observed for the primers RM263 (0.67), RM159 (0.59) and RM333 (0.50). Furthermore, out of 38 SSR markers screened on 192 temperate rice germpalsm lines, R4M17 accurately differentiated indica and temperate japonica genotypes amplifying 220 bp and 169 bp, respectively. Accordingly, 15 genotypes were reported as indica and 28 temperate japonica in addition to 149 genotypes as intermediate types. CONCLUSION: The information on marker-based diversity and performance based on cooking quality and agronomic traits helped to select the most divergent lines for crossing. Also the analysis was useful to classify the temperate germplasm into indica and temperate japonica. The classification could be helpful to devise a strategy for inter-sub species hybridization to breed for improved rice varieties.

Cardiac structural and functional abnormalities in primary hyperparathyroidism.

PURPOSE: Studies on cardiac structural and functional abnormalities in primary hyperparathyroidism (PHPT) have yielded conflicting and inconsistent results. In this prospective case-control study, we sought to compare cardiac structure and function in symptomatic PHPT patients and controls. METHODS: One hundred consecutive symptomatic PHPT patients and 113 matched controls underwent echocardiographic evaluation by the same operator. RESULTS: Left ventricular mass index (LVMI) was significantly higher in patients as compared to controls, (median of 90.95 g/m2 vs 86.5 g/m2, p = 0.041). Patients had significantly lower early trans-mitral diastolic flow (E velocity) as compared to controls (57.13 ± 14.88 vs 64.76 ± 15.45 cm/s, p < 0.001). Patients also had significantly lower early to late mitral annular velocity (E/A) as compared to controls (0.98 ± 0.37 vs 1.10 ± 0.34, p 0.013). Patients had higher frequency of aortic valve calcification (29% vs 2.65%, p < 0.001), mitral annular calcification (23% vs. 4.42%, p < 0.001), myocardial and septal calcifications (25% vs none, p < 0.001) as compared to controls. Serum PTH, calcium and uric acid significantly correlated with calcifications. Serum calcium showed a negative correlation with E/A ratio. CONCLUSIONS: Symptomatic patients with PHPT have substantial cardiac structural and functional abnormalities. These abnormalities include elevated LVMI, diastolic dysfunction, and aortic valve, mitral annular, septal and myocardial calcifications. We strongly suggest and conclude that the evaluation of PHPT patients should not only include traditional end organs like bones and kidneys but also the cardiovascular system in the form of echocardiography to detect subclinical cardiac dysfunction so that the cardiovascular health of such patients can be optimized.

Land-use systems regulate carbon geochemistry in the temperate Himalayas, India.

The Himalayan ecosystem is critical for ecological security and environmental sustainability. However, continuous deforestation is posing a serious threat to Himalayan sustainability. Changing land-use systems exert a tenacious impact on soil carbon (C) dynamics and regulate C emissions from Himalayan ecosystem. Therefore, this study was conducted to determine the changes in different C pools and associated soil properties under diverse land-use systems, viz. natural forest, natural grassland, maize field converted from the forest, plantation, and paddy field of temperate Himalaya in the surface (0-20 cm) and subsurface (20-40 cm) soils. The highest total organic carbon (24.24 g kg-1) and Walkley-black carbon contents (18.23 g kg-1), total organic carbon (45.88 Mg ha-1), and Walkley-black carbon stocks (34.50 Mg ha-1) were recorded in natural forest in surface soil (0-20 cm depth), while soil under paddy field had least total organic carbon (36.45 Mg ha-1) and Walkley-black carbon stocks (27.40 Mg ha-1) in surface soil (0-20 cm depth). The conversion of natural forest into paddy land results in 47.36% C losses. Among the cultivated land-use system, minimum C losses (29.0%) from different pools over natural forest system were reported under maize-filed converted from forest system. Land conversion causes more C losses (21.0%) in surface soil (0-20 cm depth) as compared to subsurface soil. Furthermore, conversion of forest land into paddy fields increased soil pH by 5.9% and reduced total nitrogen contents and microbial population by 28.0% and 7.0%, respectively. However, the intensity of total nitrogen and microbial population reduction was the lowest under maize fields converted from the forest system. The study suggested that the conversion of natural forest to agricultural land must be discouraged in the temperate Himalayan region. However, to feed the growing population, converted forest land can be brought under conservation effective maize-based systems to reduce C loss from the intensive land use and contribute to soil quality improvements and climate change mitigation.

Identification of Immunodominant Outer Membrane Proteins of Fusobacterium necrophorum from Severe Ovine Footrot By MALDI-TOF Mass Spectrometry.

The aim of this study was to identify the immunodominant outer membrane proteins (OMPs) of Fusobacterium necrophorum from sheep affected with severe foot-rot. The OMP profile of ovine strains of F. necrophorum has not been well studied. We analyzed the OMP profile of the most frequent lktA variant JKS-F3 of F. necrophorum associated with severe ovine foot-rot with lesion score 4 in order to identify its major immunodominant OMPs. Electrophoretic separations of extracted OMPs showed a number of spots in two-dimensional electrophoretic gels. Two immunoreactive proteins of size around 43 kDa were identified through western blotting using hyperimmune sera raised in rabbits. These two immunogenic OMPs were analyzed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF/MS) which revealed that these two OMPs of lktA variant JKS-F3 of F. necrophorum showed 46 and 42 percent protein sequence coverage and scores of 125 and 114, respectively, with the reported 43 kDa outer membrane protein of F. necrophorum strain H05, a putative porin having properties similar to pore-forming proteins of anaerobic Gram-negative bacteria. These identified immunogenic OMPs will contribute to our understanding of the pathogenic role played by this organism in ovine foot-rot and could be exploited to devise an effective control strategy through development of an OMP-based recombinant vaccine to mitigate foot-rot in sheep and goats.

Assessment of trends in climatic extremes from observational data in the Kashmir basin, NW Himalaya.

The present study aims to assess the recent changes and trends in the extreme climate indices in the Kashmir basin using the observational records from 1980 to 2016. The extreme climate indices were computed using the ClimPACT2 software and a total of 39 indices were selected for the analysis having particular utility to various sectors like agriculture, water resources, energy consumption, and human health. Besides adopting the station scale analysis, regional averages were computed for each index. In terms of the mean climatology, an increase has been observed in the annual mean temperature with a magnitude of 0.024 °C/year. Further, differential warming patterns have been observed in the mean maximum and minimum temperatures with mean maximum temperature revealing higher increases than mean minimum temperature. On the other hand, the annual precipitation shows a decrease over most of the region, and the decreases are more pronouncing in the higher altitudes. The trend analysis of the extreme indices reveals that in consonance with the rising temperature there has been an increase in the warm temperatures and decrease in the cold temperatures across the Kashmir basin. Furthermore, our analysis suggests a decrease in the extreme precipitation events. The drought indices viz., Standardised Precipitation Index (SPI), and Standardised Precipitation Evapotranspiration Index (SPEI) manifest decreasing trends with the tendency towards drier regimes implying the need for better water resource management in the region under changing climate.

Primary hyperparathyroidism and pancreatitis.

PURPOSE: The true association between primary hyperparathyroidism (PHPT) and pancreatitis continues to be controversial. In this study, we present clinical data, investigative profile, management and follow-up of PHPT patients with pancreatitis and compare this group with PHPT patients without pancreatitis. METHODS: Records of 242 patients with PHPT managed at our center over 24 years were retrospectively analyzed for demographic and laboratory data. The diagnosis of pancreatitis was entertained in the presence of at least two of the three following features: abdominal pain, levels of serum amylase greater than three times the normal or characteristic features at imaging. RESULTS: Fifteen (6.19%) of the 242 consecutive patients with PHPT had had pancreatitis. Fourteen patients (93.3%) had acute pancreatitis (AP), while one patient had chronic calcific pancreatitis. Over half (8 of 14) of the patients with AP had at least two episodes of pancreatitis. Pancreatitis was the presenting symptom in 14 (93.3%) patients. None of the pancreatitis cases had additional risk factors for pancreatitis. PHPT patients with pancreatitis had significantly higher serum calcium and ALP than PHPT patients without pancreatitis. After successful parathyroidectomy, 14 patients had no further attacks of pancreatitis during a median follow-up of 16 months (range 2-41 months), while recurrence of pancreatitis was seen in one patient. CONCLUSIONS: We conclude that pancreatitis can be the only presenting complaint of PHPT. Our study highlights the importance of fully investigating for PHPT in any pancreatitis patient with high normal or raised serum calcium level, especially in the absence of other common causes of pancreatitis.

Genotyping and antibiotic resistance patterns of Campylobacter fetus subsp.venerealis from cattle farms in India.

Bovine genital campylobacteriosis caused by Campylobacter fetus subsp. venerealis (Cfv) is of considerable economic importance to the cattle industry worldwide. Cfv causes syndrome of temporary infertility in female cattle, early embryonic mortality, aberrant oestrus cycles, delayed conception, abortions and poor calving rates. In the present study, a total of 200 samples obtained from vaginal swabs, cervicovaginal mucous (CVM), preputial washes and semen straws were investigated that were obtained from organized cattle farm of MLRI, Manasbal and unorganized sectors. Out of a total of 200 samples, 49 (47·57%) vaginal swabs, 1 (3·33%) preputial wash and 8 (25%) carried out CVM samples were positive for Cfv, whereas none of the semen straws were positive for Cfv. A total of eleven isolates of Cfv were recovered. PFGE (Pulse field gel electrophoresis) analysis revealed four different pulsotypes (I-IV) circulating in the screened farms. A common pulsotype circulating among farms could not be established. Insertion element (ISCfe1), a 233 bp amplicon of Cfv, was sequenced and the sequence was deposited in GenBank (accession no: MK475662).

Retraction of reconstructing disasters and adaptation scenario of nineteenth-century Kashmir.

The above article from Disasters, published online on 24 June 2019 in Wiley Online Library (http://wileyonlinelibrary.com) has been withdrawn by agreement between the authors, the Journal Editors, Sara Pantuliano, John Twigg, Helen Young and Matthew Foley, and John Wiley & Sons Ltd., on behalf of Overseas Development Institute. Reference Ahmad, B., Alam, A., Bhat, M. S. and Bhat, K. A. (2019), Reconstructing disasters and adaptation scenario of nineteenth-century Kashmir. Disasters. Accepted Author Manuscript. doi:10.1111/disa.12364.

Determination of prevalence, serological diversity, and virulence of Dichelobacter nodosus in ovine footrot with identification of its predominant serotype as a potential vaccine candidate in J&K, India.

The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.

Comparison of efficiency of in vitro cloned sheep embryo production by conventional somatic cell nuclear transfer and handmade cloning technique.

Conventional somatic cell nuclear transfer (SCNT) technique of in vitro production of cloned embryos involves use of costly and complicated micromanipulators. Handmade cloning (HMC) technique has been applied as efficient and cost-effective alternative in many livestock species. The aim of the present study was to compare the efficiency of in vitro production and in vitro development of cloned sheep embryos by the two techniques. Cloned embryos were produced by conventional SCNT using micromanipulator apparatus and by HMC technique. Enucleation efficiency and efficiency of fusion with somatic cell (nucleus donor) were compared. Cleavage percentage was observed on day 2 of in vitro culture (IVC), and morula and blastocyst percentages were calculated on day 7 of IVC. Higher enucleation efficiency (96.98 ± 1.01 vs. 93.62 ± 1.03; p > .05) as well as fusion efficiency was obtained with HMC technique than with conventional SCNT (96.26 ± 1.34 vs. 92.63 ± 0.70, p < .05); 181 cloned sheep embryos were produced in vitro by conventional SCNT and 92 by HMC. Cleavage percentage observed on day 2 of in vitro culture was higher in HMC than SCNT (66.92 ± 3.72 vs. 55.97 ± 2.5, respectively, p < .05). Morula percentage obtained was higher in SCNT than HMC (44.12 ± 2.93 vs. 30.43 ± 6.79, respectively, p < .05), whereas blastocyst percentage obtained by HMC was higher (12.46 ± 4.96) than SCNT (5.31 ± 2.25; p > .05). It was inferred that HMC technique provides a cost-effective and efficient method of in vitro production of cloned sheep embryos with a comparatively simpler technique with a possibility of automation. Efficiency of cloned embryo production could be improved further by propagating and standardizing this technique.

The detection and prevalence of leukotoxin gene variant strains of Fusobacterium necrophorum in footrot lesions of sheep in Kashmir, India.

The objective of this study was to determine the prevalence and identification of leukotoxin gene, lktA, variant strains of Fusobacterium necrophorum in the footrot lesions of sheep. The detection of F. necrophorum was carried out by PCR targeting the lktA gene fragment and identification of lktA variant strains was done by PCR-single stranded conformational polymorphism (PCR-SSCP) and gene sequencing. Of the 450 swabs collected from footrot lesions of sheep, 117 were lktA-positive for F. necrophorum. Of the 50 swabs collected from apparently asymptomatic sheep, only one was lktA-positive for F. necrophorum. The overall prevalence of F. necrophorum in footrot affected sheep in Kashmir valley was 26%, and ranged from 20 to 34.8%, respectively. PCR-SSCP of lktA gene fragment analysis revealed three lktA variants, designated as JKS-F1/F2/F3, while two samples (1.7%) showed multiple lktA variant strains of F. necrophorum in a single footrot-affected sheep hoof. This appears to be the first report on the presence of more than one lktA variant of F. necrophorum in a footrot lesion of sheep. The JKS-F3 lktA variant was the most frequent (75.4%), followed by JKS-F2 (14.4%) and JKS-F1 (8.4%), respectively. Among the three lktA variants identified, JKS-F3 was detected in 74 (86.0%) samples from severe footrot affected sheep with a lesion score of 4. The data suggest that JKS-F3 is the predominant lktA variant of F. necrophorum and is associated with severe footrot in sheep. Hence, JKS-F3 may be a significant variant contributing to the severity and duration of the disease in sheep.

Risk indicators for prevalence, extent and severity of periodontitis among rural Indian population aged 35-54 years.

AIMS: This study aimed to identify risk indicators associated with periodontitis and the contribution of each of the indicators towards the prevalence, extent and severity of periodontitis in a rural Indian population. METHODS: A cross-sectional study design was used to collect data according to National Survey of Adult Oral Health Australia guidelines. A multistage stratified random sampling was followed to select 1401 participants, who were in the age group of 35-54 years. The participants were selected from 50 villages belonging to the 5 sub-provinces of 2 Indian districts. Data were collected through face-to-face interviews and oral examination. Statistical analysis was performed using SAS version 9.3. The univariate, bivariate and multivariate analyses were performed to determine the risk indicators of prevalence, extent and severity of periodontitis. Population attributable fraction was estimated for each of the significant risk indicators of prevalence and extent. RESULTS: In this study, factors such as age, education, tobacco chewing and plaque accumulation were significantly associated with the prevalence of periodontitis. Age, socioeconomic status, method of tooth cleaning, alcohol consumption and plaque accumulation were significant risk indicators for generalized periodontitis. Age, tobacco chewing and plaque were associated with severity of periodontitis in the population. CONCLUSION: The rural population had a high prevalence of periodontitis. Sociodemographic factors, poor oral hygiene, tobacco and alcohol were the main risk indicators attributable to periodontitis.

Type 2 diabetes and metabolic syndrome - adipokine levels and effect of drugs.

Type 2 diabetes mellitus (T2DM) is a consequence of complex interactions among multiple genetic variants and environmental risk factors. This complex disorder is also characterized by changes in various adipokines. In this study, our objective was to estimate the levels of adiponectin, leptin, and resistin (ALR) in T2DM patients, besides studying the effect of various drugs on their levels. Study participants included 400 diabetic and 300 normal patients from the Department of Endocrinology and Department of Biochemistry, Govt Medical College Srinagar. Subjects were categorized under various groups, i.e., Group 1 (metformin treated) and Group 2 (glimepiride treated), and cases were also categorized as obese with T2DM (Group A), obese without T2DM (Group B), and T2DM only (Group C). The serum ALR levels were estimated by ELISA (Alere), and biochemical parameters were also evaluated before and after treatment. Adiponectin levels were found to be significantly lower in T2DM cases as compared to controls (12 ± 5.5 versus 22.5 ± 7.9 µg/ml), while leptin and resistin levels were found to be significantly higher than controls (14.3 ± 7.4 versus 7.36 ± 3.73 ng/ml) (13.4 ± 1.56 versus 7.236 ± 2.129 pg/ml). Taking the effect of drugs into consideration, the effect on adiponectin and resistin levels was found to be highly significant in Group 2 before and after treatment (11 ± 5 versus 19.2 ± 4.5 µg/ml) (13.6 ± 2.5 versus 7.3 ± 2.9 pg/ml), while more effect was observed in leptin among Group 1 (metformin)-treated cases (27 ± 15 ng/ml versus 15 ± 15 ng/ml). Further the adiponectin levels were found to be significantly lower in Group B, while leptin and resistin levels were found to be significantly higher among obese cases when compared to T2DM cases only. Glimepiride also shows more effect on FBG, HbA1c% levels, while metformin shows more effect on Lipid profile levels. From the study, it can be concluded that ALR levels are affected by use of antidiabetic drugs among which glimepiride shows more effect on adiponectin and resistin levels, while leptin gets affected more by metformin. It can also be proposed that ALR levels are not affected by diabetes only, suggesting that their alterations in T2DM may be due to obesity as we observed more ALR changes in obese cases when compared to T2DM cases, and so there might be an important link between adiposity and insulin resistance.

Identification and characterization of 20 novel pathogenic variants in 60 unrelated Indian patients with mucopolysaccharidoses type I and type II.

Mucopolysaccharidoses (MPS), a subgroup of lysosomal storage disorders, are caused due to deficiency of specific lysosomal enzyme involved in catabolism of glycosaminoglycans. To date more than 200 pathogenic variants in the alpha-l-iduronidase (IDUA) for MPS I and ∼500 pathogenic variants in the iduronate-2-sulphatase (IDS) for MPS II have been reported worldwide. The mutation spectrum of MPS type I and MPS type II disorders in Indian population is not characterized yet. In this study, we carried out clinical, biochemical, molecular and in silico analyses to establish the mutation spectrum of MPS I and MPS II in the Indian population. We conducted molecular analysis for 60 MPS-affected patients [MPS I (n = 30) (Hurler syndrome = 17, Hurler-Scheie syndrome = 13), and MPS II (n = 30) (severe = 18, attenuated = 12)] and identified a total of 44 [MPS I (n = 22) and MPS II (n = 22)] different pathogenic variants comprising missense, nonsense, frameshift, gross deletions and splice site variants. A total of 20 [MPS I (n = 14), and MPS II (n = 6)] novel pathogenic sequence variants were identified in our patient cohort. We found that 32% of pathogenic variants detected in IDUA were recurrent and 25% in MPS II. This is the first study revealing the mutation spectrum of MPS I and MPS II patients in the Indian population.