An associated microbiome of any host helps it in different metabolic processes ranging from the decomposition of food to the maturation of gametes. Organisms with a parasitic mode of life, though present at nutritious sites inside their host, maintain their own microbiome. Nevertheless, the comprehensive characterization and functionality of microbiome in parasitic organisms remain understudied. We selected two nematode parasites of Kashmir Merino sheep viz; Haemonchus contortus and Trichuris ovis based on their higher prevalence, difference in mode of nutrition, habitation site and effect on host. The objective of the study was to explore the bacteria associated with these parasitic nematodes of sheep. We adopted a 16S rRNA metagenomic sequencing approach to estimate and compare the bacterial communities present in these two nematode species. Nematode parasites from Kashmir Merino sheep were identified morphologically and confirmed with DNA characterization. H. contortus was dominated by phylum Proteobacteria (57%), Firmicutes (25%), Bacteroidota (15%) and Actinobacteriota (3%). Conversely, T. ovis showed Proteobacteria (78%) followed by Firmicutes (8%), Bacteroidota (8%), Actinobacteriota (1%), Fusobacteriota (1%) and other phyla (4%). This study provides a comprehensive account of the microbiome composition of H. contortus and T. ovis, both of which are highly prevalent among Kashmir Merino sheep. Additionally, T. ovis exhibited a greater bacterial diversity compared to H. contortus. Notably, these nematodes were found to harbor certain pathogenic bacteria. This study can further be carried forward in gaining insights into the complex relationship between the microbiota of a parasite and its pathogenicity, reproductive potential and host microbiome modification.

OBJECTIVES/SPECIFIC AIMS: Previous studies suggest that genetic variants in the oxytocin receptor (OXTR) may alter oxytocin dose requirement for labor induction and may increase risk for preterm labor and neurodevelopmental disorders. However, the mechanisms of actions of these variants remain unknown. The goal of this study was to functionally characterize common missense and noncoding variants in OXTR. First, we aimed to determine the effects of missense variants on two major aspects of receptor function: calcium signaling and β-arrestin recruitment. Second, we used allelic expression imbalance assays in an effort to identify regulatory single nucleotide polymorphisms (SNPs) in noncoding regions of OXTR that alter OXTR mRNA expression. METHODS/STUDY POPULATION: We used the Exome Aggregation Consortium database to identify the 12 most prevalent missense single nucleotide variants in OXTR. To determine the functional effects of these variants, we transfected human embryonic kidney cells (a common model system used to study receptor function) with wild type OXTR, variant OXTR, or empty vector control. We used the calcium-sensitive dye Fluo4 to quantify intracellular calcium flux in response to oxytocin treatment, and used bioluminescence resonance energy transfer assays to measure recruitment of the signaling partner β-arrestin to the receptor. To investigate potential effects of noncoding SNPs on OXTR mRNA expression, we quantified allele-specific expression of OXTR in human uterine tissue obtained from participants at the time of Cesarean section. We used next-generation sequencing (Illumina MiSeq) to count alleles of a reporter SNP in OXTR exon 3. RESULTS/ANTICIPATED RESULTS: Of the 12 most prevalent missense single nucleotide variants, four were predicted to be deleterious by PolyPhen variant annotation software. We anticipate that these variants will alter receptor signaling through calcium or β-arrestin pathways. We further observed that a reporter SNP in OXTR exon 3 exhibits significant allelic expression imbalance in a subset of our myometrial tissue samples, indicating that OXTR expression may be regulated by a functional SNP. Our current work focuses on discovering the functional SNPs in OXTR responsible for the pattern of allelic expression imbalance seen in mRNA. In the future, we will seek to explore the effects of these variants on uterine function by using genome editing of uterine smooth muscle cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results suggest that both missense and noncoding variants may affect OXTR expression and function. Future studies may suggest that OXTR sequencing, genotyping, or expression analysis would be useful to identify individuals likely to respond or fail to respond to safe doses of oxytocin for labor induction. Personalizing approaches for labor induction in this way would increase the safety of oxytocin and potentially reduce maternal morbidity and mortality.

Co-circulation of Chikungunya and Dengue viral infections (CHIKV and DENV) have been reported mainly due to transmission by common Aedes vector. The purpose of the study was to identify and characterise the circulating strains of CHIKV and DENV in DENV endemic region of New Delhi during 2016. CHIKV and DENV were identified in the blood samples (n = 130) collected from suspected patients by RT-PCR. CHIKV was identified in 26 of 65 samples (40%). Similarly, DENV was detected in 48 of 120 samples (40%). Co-infection with both the viruses was identified in five (9%) of the samples. Interestingly, concurrent infection with DENV, CHIKV and Plasmodium vivax was detected in two samples. CHIKV strains (n = 11) belonged to the ECSA genotype whereas DENV-3 sequences (n = eight) clustered in Genotype III by phylogenetic analysis. Selection pressure of E1 protein of CHIKV and CprM protein of DENV-3 revealed purifying selection with four and two positive sites, respectively. Four amino acids of the CHIKV were positively selected and had high entropy suggesting probable variations. Co-circulation of both viruses in DENV endemic regions warrants effective monitoring of these emerging pathogens via comprehensive surveillance for implementation of effective control measures.

The Eastern Gangetic Plain is among the world's most intensively farmed regions, where rainfed and irrigated agriculture coexist. While the region and especially Bangladesh is a major producer of rice (Oryza sativa L. ssp. indica), there is potential to further develop sustainable rice production systems. Specifically, there is scope to include a replacement crop for the short fallow between rice crops in the dominant cropping pattern of rainfed monsoon rice harvest followed by irrigated spring rice. The aim of the current research was to identify a suitable cool-season legume crop – pea (Pisum sativum L.) or lentil (Lens culinaris Medik. ssp. culinaris) – that could be grown in the brief period between rice crops. The study comprised four crop sequence experiments comparing legume cultivars differing in maturity grown in between both long and short duration rice cultivars. These experiments were done at the Bangladesh Rice Research Institute regional station at Rajshahi over three cropping cycles. This was followed by an evaluation of pea vs. fallow between rice crops on three farmers’ fields in one cropping cycle. Here it is demonstrated that green pod vegetable pea is one of the best options to intensify the rainfed monsoon rice–fallow–spring irrigated rice cropping system, notwithstanding other remunerative rabi cropping options that could displace boro rice. The inclusion of an extra crop, pea as green pod vegetable, increased farm productivity by 1·4-fold over the dominant cropping sequence (rice–fallow–rice) and farm net income by fourfold. The study highlighted the advantages in total system productivity and monetary return of crop intensification with the inclusion of a pea crop between successive rice crops instead of a fallow period.

The cropping systems of the Eastern Gangetic Plains of Bangladesh, India and Nepal are based on rice. There is a scope to intensify such systems through diversification with lentil, the most popular food legume. Two strategies were evaluated to fit lentil into the short fallow between successive monsoonal (i.e., T. aman) and pre-monsoonal (aus) or irrigated rice (boro) crop. These were early-flowering sole-cropped lentil and relay-sown lentil into rice. Firstly, 18 early-flowering lentil lines at three contrasting sowing dates were tested over two seasons on a research station at Ishurdi in Bangladesh. Secondly, relay sowing was evaluated at the same location with six early-flowering lines and two control cultivars in two seasons. It was also assessed on ten farms in Western Bangladesh, comparing relay with sole cropping over 3 years. Flowering in the early-flowering lentil lines was consistently 9–17 days earlier, than the control cultivars, but they did not achieve an economic yield (<1·0 t/ha). Relay sowing with an existing cultivar produced an economic yield of lentil, which was comparable or higher than sole-cropped lentil in all situations. The relay-sown lentil matured in sufficient time to allow the land to be prepared for the succeeding rain-fed rice crop. It was concluded that the substitution of relay-sown lentil for fallow in the monsoonal rice–fallow–rain-fed rice cropping pattern is a useful option to intensify and diversify cropping in the Eastern Gangetic Plain.

The cliff-nesting Long-billed Vulture Gyps indicus is one of four critically endangered Asian Vultures. In India, this species has declined catastrophically, but in Pakistan only small population declines have been recorded. Mortality of this species has been linked to poisoning by veterinary diclofenac, which was banned throughout south Asia in 2006. Between 2003 and 2012 we measured abundance of adult, sub-adult, juvenile, and dead vultures, and nest occupancy and productivity at the largest known Long-billed Vulture colony in Pakistan. We compared population parameters from before (2003−2006) and after (2007−2012) the ban on veterinary diclofenac. Our data and models indicate that vulture abundance, nest occupancy, and nest productivity declined 61%, 73%, and 95%, respectively, in the three years before the diclofenac ban, and then increased 1–2 years after the ban by 55%, 52%, and 95%. Furthermore, we observed 87% of total vulture mortalities prior to the diclofenac ban. Our results demonstrate for the first time since the onset of the Asian vulture crisis that the ban on veterinary diclofenac is an effective management tool for reversing Long-billed Vulture population declines.

To study the occurrence of hepatitis B virus (HBV) and hepatitis C virus (HCV) in patients with chronic liver disease (CLD) and hepatocellular carcinoma (HCC) in Pakistan, blood samples from 105 sequential patients with biopsy-proven CLD (n = 82) and HCC (n = 23) were tested for HBV and HCV markers. Of the 105, 87 (83%) had evidence of hepatitis B exposure, 58 (55%) were positive for hepatitis B surface antigen (HBsAg), 23 (22%) had hepatitis C antibodies and 25 (24%) had detectable HCV RNA. Significantly more patients with HCC had evidence of HBV exposure in the absence of HCV markers (49/82 vs. 20/23, odds ratio 4·49,95 % CI 1·17–25·16). The proportion of patients positive for HBsAg with no HCV markers was also significantly higher in the HCC group (34/82 vs. 18/23, odds ratio 5·08, 95% CI 1·59—18·96). There were more patients with only HCV markers in the CLD group than the HCC group but the difference was not statistically significant (19/82 vs. 1/23, odds ratio 6·63, 95% CI 0·93—288·01). A modified non-isotopic restriction fragment length polymorphism study on PCR products was used to investigate the epidemiology of HCV genotypes in Pakistan. Due to depletion of the initial samples, a second series of specimens collected one year afterwards was used. Fifteen out of 40 samples had amplifiable product and all were identified as type 3. A commercial serological typing method on the same samples also confirmed that type 3 was the predominant HCV genotype in Pakistan.

The leaching behaviour of a highly saline-sodic, moderately permeable, sandy-loam soil was evaluated under continuous and intermittent submergence conditions in a longterm field study in the presence of rice and subsequent wheat and sesbania crops. Leaching curves with respect to both desalinization and desodification showed that leaching efficiency was considerably higher with intermittent than with continuous submergence. The curves were useful in determining the amount of leaching water needed for a given mode of water application to reduce harmful levels of salinity and sodicity to acceptable ones. Empirical equations were determined to fit the experimental data. Their comparison with another empirical equation from published bare-field data of this site showed that leaching efficiency under crops was higher than under fallow. From the desodification leaching curve, it is concluded that in reclamation of these soils there is no need of the application of any amendment like gypsum. The soil salinity and sodicity data recorded at different growth stages and crop yields showed that leaching during the rice growing season, under intermittent submergence without previous leaching, decreased salinity and sodicity throughout the top 100 cm of the soil to levels safe for the successful cultivation of rice and subsequently the relatively deep-rooted crops of wheat and sesbania.

The consumptive use of water by cotton (G. hirsutum L.) variety 149F, sown at three dates, was calculated using Penman Et × crop factor method. The earlier the sowing date the more water evapotranspired, mainly due to differences in the length of the growing season and the amount by which it coincided with the monsoonal depression of potential evapotranspiration. An example is given of calculating the irrigation requirements of a mid-June sown cotton, accounting for average rainfall, showing that the water requirements of cotton during the early stages may be adequately met by the establishment irrigation and rains during July. Examples are cited to show that a charged profile at the beginning of the season may sustain good growth and produce satisfactory yield.

Wheat (Triticum aestivum), mustard (Brassica juncea), gram (Cicer arietinum) and barley (Hordeum vulgare) seedlings were raised in soils of different tilth created in microplots during 1981 and 1982. Amplitude in diurnal seed-zone temperature and soil moisture content increased with tilth coarseness. At low soil moisture levels, seedling emergence of wheat and mustard decreased with increasing clod size while that of gram and barley increased. These effects were not significant at high soil moisture levels. Shoot growth of all the crops was higher under coarse tilth than under fine tilth at both moisture levels. Root growth of wheat and mustard decreased with increasing clod size while that of gram and barley increased, at both moisture levels. Mustard had the highest seedling emergence at the low moisture level. Seedling emergence and shoot growth were higher under high moisture than under low moisture for all crops. Root growth varied depending upon the type of crop.

An experiment was conducted during the 1978/79 summer at the Central Cotton Research Institute, Multan-Pakistan to evaluate the crop production patterns of two cultivars of cotton (G. hirsutum L.) differing in their reaction to heat-induced sterility. Boll set in variety ST3 amounted to 7.7% of the total fruiting points, from flowers opening during the hottest months of June, July and early August, while almost all the fruiting points produced by cultivar B557 during the same period were shed and no bolls were set. Of the flowers examined during July, 83.3% in B557 contained non-dehiscent anthers against 6.7% in ST3.

The principal chemical components of milk from the Arabian camel (Camelus dromedarius) were monitored in Jordan over one year. The analyses included total solids, fat, protein, vitamins, minerals and organic acids. Large seasonal variations in total solids and fat were apparent, with maxima in mid-winter of 139 and 39·0 g/l, respectively, and minima in August of 102 and 25·0 g/l. These differences may be sufficient to alter the sensory properties of the milk, and the fat: casein ratio may need standardisation for cheesemaking. The mean values of trace elements like zinc (5·8 mg/l), iron (4·4 mg/l) and manganese (0·05 mg/l) in Jordanian camel milk could provide valuable additions to the diet of urban populations, as could the mean concentration of vitamin C (33 mg/l). The levels of organic acids were generally higher than in bovine milk and, as with all the constituents of the milk, there were discernible patterns linking concentration and season of the year.

Nodular fasciitis is an unusual benign reactive process affecting superficial and deep fascia. Its rapid growth, rich cellularity, high mitotic activity and poorly circumscribed nature result in it being easily misdiagnosed as a sarcomatous lesion. Three cases of nodular fasciitis presenting as neck lumps are reported. They were successfully treated with local excision, with no signs of recurrence following two years of follow up. This paper describes the clinical presentation and microscopic features of this rare benign lesion and it emphasizes the need for accurate histopathology and clinical suspicion, if inappropriate aggressive management is to be avoided.