Enhancing the wheat growth through micronutrients enriched biochar under salt stress

Crop irrigation with brackish water having high Na+ is a common practice in developing countries like Pakistan; hence increasing the saline-sodic soils throughout the region. Therefore, two independent studies were conducted to evaluate the impact of minerals enriched contrasting biochars (MECBs) in mitigating Na+ toxicity on soil physicochemical properties, wheat growth, and physiology. In 1st trial, an incubation experiment was conducted having 32 treatments including simple biochar (SBc), manganese enriched biochar (MnBc), zinc enriched biochar (ZnBc), and iron enriched biochar (FeBc) applied at two rates (0.1% and 0.5%) with four different salinity levels such as 10, 20, 30, and 40 dSm−1. The soil was sampled after the 8th, 16th, and 24th day of incubation. Among the four sources of biochar, MnBc (0.1%) + 40 dSm−1 and FeBc (0.5%) + 40 dSm−1 showed efficient results to tolerate salinity stress in soil by efficiently reducing the availability of Na+ (60% and 30%) when compared to SBc (0.5%) + 40 dSm−1. Then, a pot study was conducted with 20 different treatments (control, SBc, MnBc, ZnBc, and FeBc) at four different salinity levels, i.e., 0, 5, 10, and 15 dSm−1. There were four replicates and the crop was harvested after 60 days of germination. The application of brackish water (control+15 dSm−1) significantly reduced the fresh weight of root (28%) and shoot (34%) of wheat (Triticum aestivum L.) as compared to FeBc+15 dSm−1. However, the soil Na+ toxicity was significantly decreased (23%) by the application of FeBc+15 dSm−1 as compared to control+15 dSm−1. Similarly, a significant increase was observed in leaf chl. a, b and photosynthetic rate (75%, 3 and 2-folds); however, a significant decrease was observed in transpiration rate (50%) in T. aestivum L. treated with FeBc+15 dSm−1 as compared to control+15 dSm−1. Interestingly, FeBc+15 dSm−1 also played a vital role to prevent oxidative damage by controlling the shoot CMP (63%) in T. aestivum L. by efficiently triggering the activities of shoot antioxidant enzymes such as APX (30%) and CAT (52%) when compared to control+15 dSm−1. In summary, it was evaluated that FeBc showed the highest adsorption capacity of Na+ ions to regulate salinity stress by improving soil health and crop physiology

MOLECULAR CHARACTERIZATION OF INFECTIOUS BURSAL DISEASE VIRUS FROM COMMERCIAL POULTRY IN PAKISTAN

Infectious bursal disease (IBD) is an immunosuppressive disease of young, growing chickens which results in impaired growth or mortality of rearing flocks. In the current era there is a re-emergence of very virulent Infectious Bursal Disease Viruses (vvIBDV) and classical variant (cv) IBDV strains which increased the financial losses of poultry industry worldwide. Recent studies were conducted to characterize the existing vvIBDVs prevailing in Pakistan. The suspected samples were collected from the field outbreaks during the period from 2014–2017. IBDV was detected by RT-PCR. The sequences of VP2 gene (hyper variable region) were determined and available details were aligned with sequences submitted inGenBank. Phylogenetic analysis reveals that both vvIBDV and classical variant strains were circulating in different regions of Pakistan. In Indo-Pak isolates, the presence of virulent markers, amino acids (A222, I242, Q253, I256 and S299) and “Serine rich- heptapeptide” indicated the presence of very virulent viruses. The presence of T284A isan indicator of vvIBDVs in local poultry farms. More than 99% similarity of Pakistani isolates with Indian sequences reflects the trans-boundary spread of IBD. In recent studies amino acid, Glutamine (Q) is present at position 221 (as reported in previous studies) rather than Histidine (H) in Pakistani sequences. It is investigated that Glutamic acid (E) is located at position 300 in minor hydrophilic region III of VP2 protein in all reported Pakistani isolates. It is the unique feature of indigenous strains. This study will be useful in understanding the origin and pathotypes of IBDV circulating in Pakistan

A Reverse transcription-polymerase chain reaction (RT-PCR) based detection of foot and mouth disease in District Faisalabad, Pakistan during the Year 2016

Foot and mouth disease is an economically devastating disease of livestock that mainly effect cloven-hoofed animals i.e. sheep, goat, cattle, pig, buffalo, deer etc. The aim of this study was to determine the serotypes circulating in the region during 2016. Sampling was done from different outbreaks initially on the basis of clinical signs and later reverse transcriptase-polymerase chain reaction (RT-PCR) was employed for the confirmation of FMDV genome. Out of total 72 samples, 65 were found positive which were then serotyped into type O (n=30), Asia1 (n=19) and A (n=5). Some samples (n=5) were found positive for more than one serotype that were subjected to reverse transcriptase loop-mediated isothermal amplification assay (RT-LAMP) for serotype determination

COMPARISON AND EVALUATION OF ONE-STEP REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR) AND REVERSE TRANSCRIPTASE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ASSAY(RT-LAMP) FOR RAPID DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS (FMDV)

Foot-and-mouth disease (FMD) is endemic in Pakistan and cause severe economic losses. Serotype O, A and Asia 1 is prevalent and their rapid detection for Pakistaniisolates is needed to determine the serotype prevalence in different areas of the country. One step reversetranscriptase-polymerase chain reaction (RT-PCR) and reverse transcriptase loop-mediated isothermal amplification (RT- LAMP) was compared for the rapid and sensitive detection of FMDV genome. FMDV genome was confirmed in 19/25clinically affected animals by sequencing. These positive samples (n=19) were also confirmed in both RT-PCR and RT-LAMP consensus assay.In theserotyping assay, RT-PCR detection rate was16.67%, 80%, and 50% and RT-LAMP detection rate was 50%, 60% and 50%for serotype Asia 1, O and type A respectively. In general detection of FMDV, RT-PCR assay and the RT-LAMP assay showed high concordance (k = 1.0). However, in serotype detection, RT-LAMP was found more sensitive as compared to RT-PCR for the detection of the FMDV serotype Asia1 and Vice Versa in the detection of serotype O. To the best of the author’s knowledge, this is thefirst document on thecomparison of RT-LAMP and one-step RT- PCR for FMDV using ESE-Quant Tube Scanner in Pakistan.The RT-LAMP assay has the potential for early and rapid clinical diagnosis, surveillance and serotyping of FMDV infection in endemic countries

Novel VPS13B Mutations in Three Large Pakistani Cohen Syndrome Families Suggests a Baloch Variant with Autistic-Like Features.

BackgroundCohen Syndrome (COH1) is a rare autosomal recessive disorder, principally identified by ocular, neural and muscular deficits. We identified three large consanguineous Pakistani families with intellectual disability and in some cases with autistic traits.MethodsClinical assessments were performed in order to allow comparison of clinical features with other VPS13B mutations. Homozygosity mapping followed by whole exome sequencing and Sanger sequencing strategies were used to identify disease-related mutations.ResultsWe identified two novel homozygous deletion mutations in VPS13B, firstly a 1 bp deletion, NM_017890.4:c.6879delT; p.Phe2293Leufs*24, and secondly a deletion of exons 37-40, which co-segregate with affected status. In addition to COH1-related traits, autistic features were reported in a number of family members, contrasting with the "friendly" demeanour often associated with COH1. The c.6879delT mutation is present in two families from different regions of the country, but both from the Baloch sub-ethnic group, and with a shared haplotype, indicating a founder effect among the Baloch population.ConclusionWe suspect that the c.6879delT mutation may be a common cause of COH1 and similar phenotypes among the Baloch population. Additionally, most of the individuals with the c.6879delT mutation in these two families also present with autistic like traits, and suggests that this variant may lead to a distinct autistic-like COH1 subgroup

<i>Trichoderma asperellum</i> L. Coupled the Effects of Biochar to Enhance the Growth and Physiology of Contrasting Maize Cultivars under Copper and Nickel Stresses

Crop cultivation in heavy metal (HM)-polluted soils is a routine practice in developing countries that causes multiple human health consequences. Hence, two independent studies have been performed to investigate the efficiency of rice husk biochar (BC) and three fungal species, Trichoderma harzianum (F1), Trichoderma asperellum (F2) and Trichoderma viride (F3), to improve the growth and physiology of Zea mays L. plants grown on soil contaminated with Cu and Ni. Initially, a biosorption trial was conducted to test the HM removal efficiency of species F1, F2 and F3. Among them, F2 sp. showed the maximum Cu and Ni removal efficiency. Then, a pot study was conducted with two cultivars (spring corn and footer corn) having eleven treatments with three replicates. The results demonstrated a significant genotypic variation among both cultivars under applied HM stress. The maximum decreases in leaf Chl a. (53%), Chl b. (84%) and protein (63%) were reported in footer corn with applied Cu stress. The combined application of biochar and F2 increased leaf CAT (96%) in spring corn relative to Cu stress. Altogether, it was found that BC + F2 treatment showed the maximum efficiency in combatting Cu and Ni stress in spring corn

Evaluating the importance of different computed tomography scan-based factors in predicting the outcome of extracorporeal shock wave lithotripsy for renal stones

Purpose: To evaluate the impact of various computed tomography scan-based parameters of renal stones on the outcome of extracorporeal shock wave lithotripsy (ESWL). Materials and Methods: We conducted a retrospective study of patients who underwent ESWL for renal stones (sized 5–20 mm) from January 2013 to December 2016. We evaluated body mass index, location of the stone, skin-to-stone distance (SSD), stone attenuation value (SAV), stone diameter, Hounsfield density, stone area, and stone volume. Statistical analysis was done and significance was confirmed by multivariate logistic regression analysis. Results: Of the 203 patients 122 (60.1%) had successful clearance of the stone. The presence of a double J stenting, a lower pole location, a higher SAV, higher Hounsfield density, larger stone area, larger stone diameter, and higher stone volume were negative predictors of ESWL outcome. When these parameters were analyzed with multivariate logistic regression analysis, stone location, SSD, and SAV were the only significant independent predictors of the outcome of ESWL. Conclusions: Stone location, SSD, and SAV are reliable and strong predictors of ESWL outcome for the treatment of renal stones

Post-procedural Drainage versus no Drainage in Laparoscopic Cholecystectomy at DHQ Hospital Charsadda, Pakistan

Background: Laparoscopic cholecystectomy (LC) is the treatment of choice for symptomatic gall stones but controversy regarding the routine use of drainage after elective LC still exists. The objective of this study was to determine the efficacy of post-procedural drainage versus no drainage after simple laparoscopic cholecystectomy. The postoperative complications were also evaluated in both groups after the LC procedure.Material and Methods: This Randomized controlled trial (RCT) was conducted in patients who underwent Laparoscopic cholecystectomy according to a pre-set inclusion criterion. Ninety-three patients were randomly assigned into group A (with drainage tube) and group B (without drainage tube) using sealed opaque envelopes containing computer‐generated random numbers. Primary (like duration of hospital stay, Postoperative pain) and secondary outcomes (like postoperative complications) were noted in both groups. Chi-square, Fischer exact test and Mann witney U test were applied as appropriate and statistical significance was established at P &lt; .05.Results: The number of patients with hospital stay exceeding two days were more in group A (n=23; 51.1%) than B (n=13; 28.8%) (P &lt; .05). Group A presented with more postoperative complications but differences between the two groups were statistically non-significant. Both Groups experienced a high level of pain at six hours of surgery followed by progressive decrease in severity at 24 and 48 hours, respectively (P=.06).Conclusions: Post-procedural drain placement after laparoscopic cholecystectomy has no advantages as there is no significant difference in post-operative complications and duration of hospital stay in drainage versus no drainage groups

Development of amplified fragment length polymorphism (AFLP) markers for the identification of Cholistani cattle

&lt;p&gt;The identification issue of livestock can be resolved by using molecular identification tools that are acceptable to preserve and maintain pure breeds worldwide. The application of a molecular identification methodology is more important for developing nations, e.g., Pakistan, where uncontrolled crossbreeding has become a common practice and the import of exotic animals and germplasm is ever increasing. This presents a risk to local breeds as also stated by the FAO. Therefore, the current study was designed to develop standard molecular markers for Cholistani cattle to ascertain their purity for breeding purpose. In this study 50 and 48 unrelated males were sampled for Cholistani and each crossbred cattle, respectively. Candidate molecular markers present in Cholistani but absent in crossbred cattle and vice versa were detected using the amplified fragment length polymorphism (AFLP) method. Eleven markers were developed and were converted to single nucleotide polymorphism (SNP) markers for genotyping. The allele frequencies in both breeds were determined for discrimination ability using polymerase-chain-reaction–restriction-fragment-polymorphism (PCR-AFLP). The probability of identifying the Cholistani breed was 0.905 and the probability of misjudgment was 0.073 using a panel of markers. The identified markers can ascertain the breed purity and are likely to extend the facility for breed purity testing before entering into a genetic improvement program in the country.&lt;/p&gt