Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Australian Uromyces viciae-fabae: Host and nonhost interaction among cultivated grain legumes

Uromyces viciae-fabae, rust of faba bean, parasitizes other legume crops such as lentils (Lens culinaris) and field peas (Pisum sativum) in some environments. In this study we examined the host range of two Australian isolates of U. viciae-fabae collected and purified from a faba bean crop and classified as U. viciae-fabae ex V. faba. Field pea (P. sativum), chickpea (Cicer arientinum), lupin (Lupinus spp.), lentil (L. culinaris), and mung bean (Vigna radiata) genotypes were tested with these isolates, as well as resistant and susceptible genotypes of the faba bean host. Race specificity for these two pathogen isolates was observed on Vicia faba, with two faba bean genotypes showing partial resistance. Both U. viciae-fabae isolates also colonized field pea seedlings and successfully produced uredinia under glasshouse conditions, despite this fungus not being known as a pathogen of Australian field pea crops. No sporulation of either isolate of U. viciae-fabae ex V. faba was observed on any of the remaining legume species tested. However, obvious differences in fungal growth were observed, ranging from small infection sites with very rare haustorium formation in mung bean to more extensive growth and the development of potential uredinial structures in chickpea. These observations are discussed in relation to the phylogenetic relationship of these host and nonhost species.     

Dynamics of organic carbon and nitrogen in deep soil profile and crop yields under long-term fertilization in wheat-maize cropping system

Soil organic carbon (SOC) and nitrogen (N) are two of the most important indicators for agricultural productivity. The primary objective of this study was to investigate the changes in SOC and N in the deep soil profile (up to 100 cm) and their relationships with crop productivity under the influence of long-term (since 1990) fertilization in the wheat-maize cropping system. Treatments included CK (control), NP (inorganic N and phosphorus (P) fertilizers), NPK (inorganic N, P and potassium fertilizers), NPKM (NPK plus manure), and M (manure). Crop yield and the properties of topsoil were measured yearly from 2001 to 2009. C and N contents were measured at five different depths in 2001 and 2009. The results showed that wheat and maize yields decreased between 2001 and 2009 under the inorganic fertilizer (NP and NPK) treatments. The average yield between 2001 and 2009 under the NP, NPK, NPKM, and M treatments (compared with the CK treatment) increased by 38, 115, 383, and 381%, respectively, for wheat and 348, 891, 2 738, and 1 845%, respectively, for maize. Different long-term fertilization treatments significantly changed coarse free particulate (cfPOC), fine free particulate (ffPOC), intramicroaggregate particulate (iPOC), and mineral-associated (mSOC) organic carbon fractions. In the experimental years of 2001 and 2009, soil fractions occurred in the following order for all treatments: mSOC>cfPOC>iPOC>ffPOC. All fractions were higher under the manure application treatments than under the inorganic fertilization treatments. Compared to the inorganic fertilization treatments, manure input enhanced the stocks of SOC and total N in the surface layer (0–20 cm) but decreased SOC and N in the deep soil layer (80–100 cm). This reveals the efficiency of manure in increasing yield productivity and decreasing risk of vertical loss of nutrients, especially N, compared to inorganic fertilization treatments. The findings provide opportunities for understanding deep soil C and N dynamics, which could help mitigate climate change impact on agricultural production and maintain soil health.     

Follicular dynamics and changes in oestradiol‐17β, progesterone and LH profiles following PGF2α induced oestrus in early and late ovulating Beetal goats

This study compares the factors associated with variable interval to oestrus and ovulation between early versus late ovulating goats following PGF_2α administration. The time of ovulation in Beetal goats (n = 38) was monitored through transrectal ultrasound at every 6 hr following a single dose of PGF_2α (experiment 1). Variations in oestrus and ovulation times were further explored through the changes in follicular dynamics, endocrine profiles and behaviour in another set of goats (n = 13) following single PGF_2α given randomly during the luteal phase (experiment 2). The ovulation time varied between 60 and 96 hr, and 57% of ovulations occurred by 72 hr following PGF_2α (experiment 1). Accordingly, the goats (n = 13) in the second experiment were retrospectively divided either into early and/or late ovulating, that is, ≤72 and/or ≥84 hr following PGF_2α. The onset of oestrus, peak estradiol‐17β concentration and LH surge after PGF_2α was first observed in early than late ovulating goats (p < 0.05). The goats ovulating early had larger follicle and smaller CL in diameter at the time of PGF_2α administration than those ovulating late (5.4 ± 0.2 vs. 4.3 ± 0.2 mm and 10 ± 0.6 vs. 11.8 ± 0.3 mm, respectively; p < 0.05). Likewise, plasma progesterone concentration tended to be lower (p = 0.087) in early than late ovulating goats. In conclusion, the size of dominant follicle and CL at the time of PGF_2a determines the interval to ovulation following a single dose of PGF_2a during the luteal phase.     

The first report on serotyping of bluetongue virus in small ruminants of Khyber Pakhtunkhwa province,Pakistan

Bluetongue virus (BTV), a member of Orbivirus genus (family Reoviridae), is a non-contagious infection of domestic and wild ruminants. The current study was designed to detect various serotypes of BTV in small ruminants of Khyber Pakhtunkhwa (KPK) province of Pakistan, along with their effects on hemato-biochemical parameters. A total of 408 serum samples in four districts (Mansehra, Abbottabad, Swabi, and Kohat) of KPK from small ruminants were screened based on competitive ELISA (cELISA). A total of 204 (50%) samples were found positive for BTV group–specific antibodies. The seropositive samples were processed for the detection of BTV serotypes through real-time polymerase chain reaction (qPCR). Out of 204 cELISA-positive samples, 60 (29.41%) were found positive through qPCR. Three serotypes [6, 8, 9] were detected from Mansehra District and two from Kohat [2, 8] and Abbottabad [6, 8], while only one from Swabi [8]. The serotype “8” was found consistently in all the four study districts. A significant (p?<?0.05) increase in the level of blood urea nitrogen (BUN) and alkaline phosphatase (ALP) was recorded in goats, whereas aspartate aminotransferase (AST) in sheep infected with BTV, compared to healthy animals. The hematological parameters showed significantly (p?<?0.05) raised total leucocyte count (TLC) in both sheep and goats, whereas only hematocrit (HCT) value was increased significantly (p?<?0.05) in infected sheep. This is the first report on serotyping of BTV among small ruminants in Pakistan.

     

Synthesis and Characterization of Aqueous Chitosan-polyurethanes Dispersion for Textile Applications with Multipurpose Performance Profile

As the use of high performance textiles has grown, the need for chemical finishes to provide the fabric properties required in the special applications has grown accordingly. In this project, a series of water dispersible polyurethanes dispersion (CS-PUs) with multipurpose performance profile was developed using isophorone diisocyanate (IPDI), polyethylene glycol (PEG), 2,2-dimethylol propionic acid (DMPA) and chitosan (CS) for textile applications. In two step synthesis process, NCO functional PU prepolymers prepared by reacting IPDI, PEG, and DMPA were extended with varying molar quantities of chitosan followed by structural characterization through FTIR. The prepared CS-PU dispersions were applied onto the dyed and printed poly-cotton blend fabrics. The performance behavior of the treated fabric in terms of crease recovery, tear strength, tensile strength, and antibacterial properties was evaluated by applying standard test methods. These investigations show that the CS-PU dispersions can be applied as antibacterial textile finishes with significant improvement in the physical and mechanical properties of poly-cotton fabrics.     

Effects of interactions among Metarhizium anisopliae, Bacillus thuringiensis and chlorantraniliprole on the mortality and pupation of six geographically distinct Helicoverpa armigera field populations

A local isolate of Metarhizium anisopliae (Hypocreales: Clavicipitaceae), Bacillus thuringiensis subsp. kurstaki and chlorantraniliprole were assessed against six field populations of tomato fruitworm Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in a series of laboratory bioassays. Two dose rates of B. thuringiensis (0.5, 1 μg g^?1), one of both M. anisopliae (1.3?×?10⁶ conidia ml^?1) and chlorantraniliprole (0.01 ppm) were applied alone and in combination with each other against 2nd, 3rd, 4th and 5th larval instars. The mortality was observed every 24 h until pupation. The bioassays were carried out at 25°C and 75% r.h. The highest mortality was observed in Rawalpindi with the lowest pupation rate by applying the combined concentrations of B. thuringiensis and chlorantraniliprole. The lowest mortality was observed in population from Gujranwala among all the tested populations. The antagonistic interaction was noted where the high dose rate of B. thuringiensis was combined with M. anisopliae; however, the remaining interactions enhanced the mortality and reduced the percent pupation. The overall results demonstrated that all the treatments gave significant control of the larval instars of H. armigera. The population from Gujranwala proved least susceptible whereas the one from Rawalpindi was highly susceptible.     

Effect of animal age,postmortem chilling rate,and aging time on meat quality attributes of water buffalo and humped cattle bulls

The study was aimed to investigate the influence of animal age, post‐slaughter chilling rate, and aging time on meat quality of M. longissimus dorsi (LD) of water buffalo (Bubalus bubalis) and humped cattle (Bos taurus indicus) bulls. After slaughtering, one side of carcasses was subjected to rapid chilling (RC) (0 ± 2°C) and other side was hanged in controlled room temperature (25 ± 2°C) for 3 hr, then allowed to the chiller (0 ± 2°C). The meat quality traits were analyzed at 1, 7, and 14 days of storage. It was noted that rapidly chilled carcasses from the younger animals of both species missed the ideal pH/temperature window, which affects the toughness of the meat. Buffalo meat presented higher shear force, color L* values, and lower b* value as compared to the cattle meat. Moreover, meat shear force values decreased while all color coordinates and cooking loss values increased with lengthening the storage time in both age groups of cattle and buffalo. In conclusion, the tenderness of cattle meat was superior to that of buffalo and RC adversely affect the shear force values of young cattle and both age groups of buffalo bulls.     

Seasonality and photoperiod influence in vitro production of buffalo embryos

Buffalo is considered short-day breeder in tropical and subtropical part of the world and seasonality and photoperiodism impart major influence on its fertility. However, its impact on in vitro embryo production (IVEP) remains elusive. Therefore, this study investigated the effect of seasonal variations and photoperiodism on morphological and molecular parameters of IVEP in buffalo. For this purpose, we conducted two different experiments on the oocytes obtained by aspirating follicles from abattoir derived ovaries. In Exp. I, retrospective analysis was performed for oocyte recovery, blastocyst and hatching rate, during four consecutive seasonal periods (i.e. January–March, April–June, July–September and October–December). In Exp. II, oocytes from peak breeding and non-breeding seasons were subjected to 24 hr in vitro maturation and evaluated for polar body extrusion to assess maturation rate. Results showed that embryo development was markedly low during second quarter (April–June) and maximum during fourth quarter (October–December) of the year; referred as non-breeding and breeding seasons, respectively. Comparative data analysis demonstrated that poor oocyte quality is major reason for lesser efficiency of embryo production during non-breeding season than peak breeding season as suggested by poor oocyte recovery (2.31 ± 0.10 vs. 3.65 ± 0.27) and maturation rate (33.32 ± 2.1 vs. 63.15 ± 7.31). Subsequently, comparative gene expression analysis of blastocysts during peak breeding season significantly upregulated pluripotency gene (OCT-4) and downregulated heat shock protein 90, as compared to non-breeding season. Therefore, it could be divulged from the present study that seasonal variations and photoperiodism have profound effect on oocyte quality and subsequent embryo development. It is recommended to find suitable additives for in vitro maturation that could mitigate seasonal effects.     

Fungal endophyte Penicillium janthinellum LK5 can reduce cadmium toxicity in Solanum lycopersicum (Sitiens and Rhe)

We investigated the role of gibberellins-producing endophyte Penicillium janthinellum LK5 associated with Solanum lycopersicum (host), abscisic acid (ABA)-deficient tomato mutant Sitiens and its wild-type Rheinlands Ruhm (Rhe) plants under cadmium (Cd) stress. A 100-μM Cd application to host, Sitiens and Rhe reduced the shoot growth, chlorophyll content and stomatal conductance. However, these parameters were significantly (P?<?0.0011) higher (1.0- to 2.6-folds) in host, Sitiens and Rhe under endophytic association than in non-endophyte infected plants (control) under Cd stress. Furthermore, endophytic association minimized the Cd-induced membrane injury and oxidative stress to host, Sitiens and Rhe plants by reducing electrolytes and lipid peroxidation while increasing the content of reduced glutathione and catalase activities as compared to non-endophyte-infected plants. Stress-responsive ABA content significantly increased (～2-folds) in Sitiens and Rhe under endophyte association, while in host plants it was decreased under Cd stress. Salicylic acid content was ～?1.7-fold higher in host, Sitiens and Rhe plants under Cd stress and endophyte association than in the control. Besides gibberellins production, the endophyte has the potential to solubilize phosphates (12.73?±?0.24 mg/l) since higher P was observed in the roots of Sitiens, Rhe and host plants. Similarly, nutrients like sulfur and calcium were more efficiently assimilated in roots of endophyte-associated plants than control under Cd stress. Conversely, Cd accumulation was significantly decreased (P?<?0.001) in the roots of endophyte-inoculated host, Sitiens and Rhe than control. In conclusion, endophyte symbiosis can counteract heavy metal stress which can exert negative effects on plant growth.