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.     

Micropropagation of Prunus scoparia,a Suitable Rootstock for Almond under Drought Conditions

Prunus scoparia is a wild deciduous shrub, usually living on dry calcareous soils of the rocky mountains and has been used as a grafting rootstock for domesticated almonds to provide drought resistance. In the current study, micropropagation ability of P. scoparia was investigated using cytokinin and auxin. Uniform nodal shoot pieces (3–5 cm in length) of seedlings were used as explants. The explants were disinfected with 10% sodium hypochlorite solution. For adventitious shoot induction and proliferation, Murashige and Skoog (MS) media containing 7.00 g/l agar and 30.00 g/l sucrose containing five concentrations of benzyl adenine (BA) (0.00, 0.50, 1.00, 2.00, and 4.00 mg/1) and also containing six concentrations of Thidiazuron (TDZ) (0.00, 0.50, 1.00, 2.00, 5.00, and 7.00 mg/1) were compared. For rooting, in vitro shoots (2–3 cm) were transferred into ½ MS medium supplemented with 30 g/l sucrose, 7.50 g/l agar, and different concentrations of IBA (0.00, 0.25, 0.50, and 1.00 mg/l) and NAA (0.00, 0.25, 0.50, and 1.00 mg/l). Based on the results obtained for shoot proliferation, only 2.00 and 4.00 mg/l BA and 2.00 mg/l TDZ concentrations generated shoots, while other treatments did not show shoot proliferation. Among the three treatments that generated shoots, the best results for shoot number, leaf number, and leaf color quality were observed in media containing 2.00 mg/l TDZ. Based on the results obtained for rooting, the effect of IBA concentrations on the rooting percentage, root number, and root length was significant. Among IBA concentrations, only 0.50 mg/l IBA induced rooting, while there was no rooting in the media containing other IBA concentrations. None of the NAA concentrations showed rooting. In conclusion, MS culture medium supplemented with 2.00 mg/l TDZ and ½ MS culture medium supplemented with 0.50 mg/l IBA are suggested for in vitro shoot proliferation and rooting of P. scoparia, respectively. The results presented herein could be used for in vitro selection and micropropagation of P. scoparia.     

Effect of thermal stress on Hsp70 gene expression and female reproductive performance of giant freshwater prawn,Macrobrachium rosenbergii

Using Hsp70 as a biomarker, thermal stress impinges on reproductive organs, ovary and hepatopancreas were being analyzed by determining the expression of Hsp70 mRNA inside the organs after the adult inter‐molt females were subjected to thermal treatment at 35, 30 and 28°C (Control). Results showed the expression of Hsp70 mRNA under thermal treatment of 35°C after 2 hr recovery in ovary were upregulated at 2, 4, 6, 12, 24 hr and 30 days compared to control whereas in hepatopancreas under similar treatment, the expression of Hsp70 mRNA were significantly higher than control at 6, 24 hr and 30 days. Frequency of reproductive molt at 35°C showed the ovary of females were failed to develop and only entered common molt along three consecutive molt cycles. For 30°C thermal treatment, the expression of Hsp70 mRNA was significantly higher than control after 2 hr recovery but returned to normal afterwards until 30 days’ thermal treatment. Maternal heat shock for 2 hr at 35°C were found to give significantly lower frequency of reproductive molt and longer duration of ovarian development and incubation period whereas maternal heat shock for 2 hr at 30°C gave lower frequency of reproductive molt, slower development of embryo and lower hatching success compared to untreated control. This study suggests that short and long‐term thermal stress at 30 and 35°C were found to affect the induction of Hsp70 mRNA in reproductive organs of Macrobrachium rosenbergii and also influence their reproductive performance.     

Dietary simultaneous replacement of fish meal and fish oil with blends of plant proteins and vegetable oils in yellowfin seabream (Acanthopagrus latus) fry: Growth,digestive enzymes,antioxidant status and skin mucosal immunity

A 56‐day nutritional research was performed to examine the influence of alternative vegetal protein and lipid sources on performance of yellowfin seabream fry (Acanthopagrus latus) (0.5 ± 0.0 g). In this regard, five isoproteic (Ca. 500 g/kg) and isolipidic (Ca. 150 g/kg) diets were formulated in which fish meal (FM) and fish oil (FO) were simultaneously replaced with blends of plant proteins (PP, soybean meal and corn gluten) and vegetal oils (VO, canola and soybean oils) at 20% (SR20), 40% (SR40), 60% (SR40) and 80% (SR80) levels, respectively; meanwhile, a control diet (SR0) was formulated based on FM and FO. Growth and feed utilization were not influenced by experimental diets. The fatty acid profile of fillet drastically altered by dietary treatments. Fish fed with the SR60 and SR80 feeds had higher total protease, trypsin and α‐amylase activities than other treatments. The antioxidant enzyme activities and glutathione content in liver were enhanced in fish fed with the SR40, SR60 and SR80 diets. Skin mucosal immune parameters including total protein content, alkaline phosphatase and alternative complement pathway activities in the control group were relatively lower than the vegetal treatments. According to these results, it is recommended that 410 g/kg of FM and 45 g of FO/kg can be replaced with alternative vegetal sources in diet for A. latus fry.     

Meat and Bone Meal as Partial Substitute for Fish Meal in Nursery Diet for Giant Freshwater Prawn, Macrobrachium rosenbergii (de Man)

An experiment was conducted for 60 d in a recirculatory system consisting of 20 glass aquariums (each of size 41 × 41 × 46 cm) to evaluate meat and bone meal (MBM) as partial replacement for fish meal (FM) in Macrobrachium rosenbergii postlarvae (PLs) with a view to develop an inexpensive nursery diet. Three nursery diets were formulated to contain 32% protein and to be as isoenergetic as possible. FM in these diets was progressively replaced with MBM (Diet 1 contained 25% FM and 14% MBM, Diet 2 contained 20% FM and 20% MBM, and Diet 3 contained 15% FM and 26% MBM). Inclusion levels of mustard oilcake and sesame meal in diets were kept fixed at 10%. A commercial shrimp nursery diet (30% protein) from Saudi Bangla Fish Feed Ltd., Mymensingh, was used as the reference diet (Diet 4). Each treatment had three replicates. PLs of M. rosenbergii (26 ± 0.02 mg) were stocked at the rate of 60 PLs (1 PL/L) in each aquarium. At the beginning, PLs were fed three times daily at a rate of 20% of the body weight and it was gradually reduced to 10% at Day 30. The ranges of water quality parameters in the system were as follows: temperature, 27–29 C; dissolved oxygen, 6.5–8.4 mg/L; pH, 6.8–8.3; and total ammonia, 0.01–0.15 mg/L. The result of the study showed that there was no significant difference (P > 0.05) between the weight gains of PLs fed Diet 1 (containing 14% MBM) and Diet 4 (reference diet) but was significantly higher than other dietary groups containing higher levels of MBM. Diets 1 and 4 showed significantly lower (P < 0.05) apparent food conversion ratios (1.74 and 1.73, respectively). The survival (%) of PLs ranged between 62 and 76% with PLs fed Diets 1 and 4 showing significantly higher (P < 0.05) survival. The result of the study demonstrated that the MBM could be included at 14% level in a nursery diet, and a diet containing 25% FM, 14% MBM, 10% sesame meal, and 10% mustard oilcake may be recommended for profitable production of PLs of M. rosenbergii.     

Characteristics of the organic matter in a slightly and in a severely crusted soil

Chemical and spectroscopic characteristics were determined on the organic matter in two Luvisolic soils from northwestern Alberta, Canada. One of these (the Debolt soil) is known to crust only slightly while the other (the Demmitt soil) exhibits severe crusting after heavy rainfalls. The Debolt top soil is richer in total C, total N, carbohydrates and proteinaceous materials than the Demmitt top soil. Humic acid extracted from the Debolt soil is more aliphatic, and contains fewer CO₂H groups than does humic acid extracted from the Demmitt soil. The quality of the organic matter appears to play a role in soil crusting.     

Evaluation of furrow fertigation and model validation on maize field

In new agricultural practices, joint application of water and fertilizer has been become common. Uniform distribution of fertilizer in soil and in plant growth duration is possible by this procedure. The main objective of this study was to investigate furrow fertigation management effects on distribution uniformity and runoff losses of nitrate in a cornfield, and validate a numerical fertigation model. A field experiment was carried out with seed corn at 12 experiments with a complete randomized block design during 2 years. Nitrogen requirement was determined by soil analysis and accomplished in four stages of the growth: before cultivation, in seven leaves, shooting and earring stages which first section (before cultivation) was applied by manual distribution and others by fertigation. Potash and super phosphate fertilizers (based on soil analysis) were sprayed on soil before planting. Water requirement was estimated by using class a evaporation pan multiplied by plant (Kc) and pan coefficients. Nitrogen fertilizer was solved in irrigation water and injected at the last minutes of irrigation. The results showed that fertilizer distribution uniformity of the low quarter (DU_LQ) ranged from 85.7% to 91.5% in first year, and 69.9% to 95.5% in second year. While water DU_LQ ranged from 74.1 to 98.2% in 2 years. Nitrate losses of surface runoff have ranged between 5.7 and 42.0% in first year according to the application time and the outflow flux. In second year, the nitrate losses decreased by adopting appropriate management based on the experiences of first year. The fertigation model was subsequently applied to the experimental data and results showed good agreement with field data.     

Physico-chemical characteristics of apricot (Prunus armeniaca L.) grown in Northern Areas of Pakistan

In order to ascertain physico-chemical, functional and geometrical traits of apricot fruit from Northern Areas of Pakistan, six predominantly grown varieties namely, Alman, Habi, Khakhas, Mirmalik, Neeli and Shai were selected in this study. Proximate composition as crude fat (2.1–3%), crude protein (6.18–8.7%), crude fiber (11.85–13.6%), ash (9.45–12.1%) and total sugars (56.8–64.9%) were determined on dry weight basis. The data showed variations among the investigated parameters in all varieties. Functional properties of apricot fruit viz. ascorbic acid (67.39–90.94 mg/100 g), total phenolic compounds (4590–7310 mgGAE/100 g), total carotenoids (10.09–18.13 mg/100 g β-carotene) and antioxidant activity (56.84–82.33%) were also recorded. The data pertaining to mineral contents (mg/100 g) revealed K as the predominant element (2040–3000) followed by P, Mg, Ca, Na and Fe among all the tested samples. Furthermore, geometrical characters of apricot varieties were also determined as important sensory and technological attributes on fresh weight basis. The result from the present study showed that all the tested varieties are highly nutritious and rich in functional components.     

Relationship between in vitro production of auxins by rhizobacteria and their growth-promoting activities in Brassica juncea L.

Rhizobacteria were isolated from the rhizosphere of different Brassica species and assayed for their ability to produce auxins in vitro. The isolates varied greatly in their potential for auxin production (ranging from 0.33 to 11.40 µg ml^-1). L-Tryptophan (an auxin precursor) addition to the media increased the auxin production by several fold. Based upon in vitro auxin production and growth promotion of B. juncea seedlings caused by various isolates under gnotobiotic conditions, promising isolates were selected and tested in pot trial to observe their effects on growth, yield and oil content of the same Brassica species. Results showed that seed inoculation with different isolates of rhizobacteria significantly increased plant height (up to 56.5%), stem diameter (up to 11.0%), number of branches (up to 35.7%), number of pods per plant (up to 26.7%), 1,000-grain weight (up to 33.9%), grain yield (up to 45.4%) and oil content (up to 5.6%) over the uninoculated control. Isolate S54 gave the most promising and consistent results. Highly significant correlations between L-TRP-derived auxin production by plant growth-promoting rhizobacteria (PGPR) in vitro and grain yield (r =0.77^**), number of pods (r =0.78^**) and number of branches per plant (r =0.77^**) of B. juncea were found. It was hypothesized that these PGPR may influence the growth and yield of inoculated plants by production of auxins in the rhizosphere of inoculated plants from the L-TRP present in the root exudates, although other mechanisms of action might have also contributed.