Prevalence and associated risk factors of ixodid tick species infesting cattle and sheep in Balochistan, Pakistan.

Tick infestation poses a serious threat to animal health, leading to significant losses in terms of vector-borne disease transmission, reduced live weight, lower quality hides, decreased milk production, and impaired reproduction in tropical and subtropical regions worldwide. This study aimed to determine the prevalence, seasonal variation, distribution pattern, and associated risk factors of Ixodid family tick species in the cattle and sheep population of three different districts in Balochistan, Pakistan. This study employed a convenient sampling method, collecting 4080 adult ticks from 816 cattle and sheep of various breeds, ages, and sexes. Specific morphological keys were used to identify the ticks up to the genus and species level. Among cattle, the highest prevalence was recorded for R. (B) annulatus (27.01%), followed by R. (B) microplus (24.02%), and H. anatolicum (20.54%). H. dromedarii (5.29%) was the least prevalent species observed in cattle. In the sheep population, H. anatolicum (30.34%) showed the highest prevalence, followed by H. marginatium (22.99%), and R. (B) annulatus (20.88%). H. dromedarii (6.96%) was the least prevalent species observed in sheep. The prevalence of R. (B) decoloratus, H. anatolicum and H. dromedarii was found to be significantly associated (P < 0.05) with the breed, age, and sex of both cattle and sheep. However, the presence of R. (B) annulatus, R. (B) microplus and H. marginatium tick species showed no significant association (P > 0.05) with these factors. In addition, the prevalence of ticks was higher in younger, female, and crossbred Friesian cattle compared to adults, males, and other breeds. Conversely, the prevalence of ticks was higher in adult, female and Hernai breed of sheep in the studied area. In conclusion, R. (B) annulatus and H. anatolicum are the dominant tick species infesting the cattle and sheep population in Balochistan. Consequently, this study provides valuable insights for developing practical and effective control measures against ticks and tick-borne diseases in the sheep and cattle population of Balochistan, Pakistan.

Molecular detection and assessment of risk factors for Theileria lestoquardi in sheep from Balochistan, Pakistan.

This study aimed to determine the molecular prevalence and associated risk factors of theileriosis in sheep from Balochistan, Pakistan. For this purpose, a total of 408 blood samples were collected from tick-infested sheep in three different zones of Balochistan (i.e., Quetta, Zhob, and Loralai). All the collected samples were analyzed using conventional microscopy techniques, polymerase chain reaction (PCR), 18S small subunit rRNA gene sequencing, and phylogenetic analysis. The results of the microscopy and PCR confirmed the highest prevalence of Theileria species in district Zhob (14.22% and 15.68%) followed by district Loralai (11.52% and 13.97%) and district Quetta (10.29% and 12.00%), respectively. In addition, the prevalence of T. lestoquardi was higher in female sheep (84.12%), followed by adult sheep (74.71%) and the Hernai breed of sheep (28.23%) in the studied area. Similarly, the prevalence of theileriosis was higher in the summer season (40.59%), followed by the spring, autumn, and winter seasons. However, numerous risk factors such as age, sex, area, season, and breeds of the sheep were not significantly correlated (P > 0.05) with the presence of T. lestoquardi, except tick abundance and feeding pattern of animals (P < 0.05). Furthermore, sequencing and phylogenetic analyses of the isolated T. lestoquardi displayed 99% sequence similarity with isolates from Germany, Egypt, Iraq, India, Iran, and Pakistan. Altogether these results showed that T. lestoquardi is the main species causing ovine theileriosis in Balochistan. As a result, large-scale studies are required to design practical control approaches to reduce the risk of theileriosis infection in Balochistan, Pakistan.

Effect of ethyl methanesulfonate mediated mutation for enhancing morpho-physio-biochemical and yield contributing traits of fragrant rice.

Background: Chemical mutagenesis has been successfully used for increasing genetic diversity in crop plants. More than 800 novel mutant types of rice (Oryza sativa L.) have been developed through the successful application of numerous mutagenic agents. Among a wide variety of chemical mutagens, ethyl-methane-sulfonate (EMS) is the alkylating agent that is most commonly employed in crop plants because it frequently induces nucleotide substitutions as detected in numerous genomes. Methods: In this study, seeds of the widely consumed Basmati rice variety (Super Basmati, Oryza sativa L.) were treated with EMS at concentrations of 0.25%, 0.50%, 0.75%, 1.0%, and 1.25% to broaden its narrow genetic base. Results: Sensitivity to a chemical mutagen such as ethyl methanesulfonate (EMS) was determined in the M1 generation. Results in M1 generation revealed that as the levels of applied EMS increased, there was a significant reduction in the germination percent, root length, shoot length, plant height, productive tillers, panicle length, sterile spikelet, total spikelet, and fertility percent as compared to the control under field conditions. All the aforementioned parameters decreased but there was an increase in EMS mutagens in an approximately linear fashion. Furthermore, there was no germination at 1.25% of EMS treatment for seed germination. A 50% germination was recorded between 0.50% and 0.75% EMS treatments. After germination, the subsequent parameters, viz. root length and shoot length had LD50 between 05.0% and 0.75% EMS dose levels. Significant variation was noticed in the photosynthetic and water related attributes of fragrant rice. The linear increase in the enzymatic attributes was noticed by the EMS mediated treatments. After the establishment of the plants in the M1 generation in the field, it was observed that LD50 for fertility percentage was at EMS 1.0% level, for the rice variety. Conclusion: Hence, it is concluded that for creating genetic variability in the rice variety (Super Basmati), EMS doses from 0.5% to 0.75% are the most efficient, and effective.

DOCUMENTATION OF TRYPANOSOMA EVANSI IN CAPTIVE TIGERS AND LIONS IN PUNJAB (2016-2018), PAKISTAN.

Trypanosoma evansi is an important hemoparasite of a variety of animal species worldwide. This parasite is a threat to the health of domestic animals as well as wild animals, particularly those managed in captivity. The current study investigated the presence of T. evansi in captive tigers (Panthera tigris tigris) and lions (Panthera leo) in Pakistan. In total, 24 blood samples from 11 tigers and 3 lions (n = 14) were collected during the course of roughly 3 yr (2016-2018). Eighteen samples were subjected to both microscopic and molecular evaluation for the presence of T. evansi; the remaining 6 samples were processed for PCR only. Of the 18 samples tested by both methods, 3 (16%) and 8 (44%) were positive by microscopy and PCR, respectively. This highlights the higher sensitivity of PCR over microscopy for detection of trypanosomes. Of the 24 total samples evaluated by PCR, 12 (50%) were positive. The three sequences obtained showed 99% identity with variant surface glycoprotein genes of the different isolates of T. evansi. The sensitivity, specificity, positive predictive value, and negative predictive value of microscopy in identifying T. evansi was 37.5, 100, 100, and 66.7%, respectively, considering PCR as the gold standard. We recommend rigorous monitoring of captive tigers and lions for hemoparasites, particularly in winter and early spring in areas with high infection rate of this parasite, preferably via PCR.

Poloxamer and Chitosan-Based In Situ Gels Loaded with Orthosiphon stamineus Benth. Extracts Containing Rosmarinic Acid for the Treatment of Ocular Infections.

Objectives: Orthosiphon stamineus Benth. (OS) is a commonly used medicinal plant for curbing bacterial infections globally. This work aimed to fabricate poloxamer and chitosan-based in situ gels loaded with standardized aqueous-ethanolic OS leaf extracts and investigate their antimicrobial efficacy as a potential remedy against ocular infections. Materials and Methods: In situ gels containing 0.5% w/v OS extract prepared using cold dispersion method were subjected to physicochemical characterization, including in vitro-release studies. Antimicrobial efficacy was tested against Staphylococcus aureus, Micrococcus luteus, Escherichia coli, and Pseudomonas aeruginosa using agar diffusion method. Results: Thin layer chromatography and high performance liquid chromatography chromatograms confirmed the presence of rosmarinic acid (RA) and sinensitin in OS extracts with same retention factor (0.26 and 0.49) and retention times (12.2 and 20.7 min) against reference standards. A homogenous brown coloured in situ gel exhibited low viscosity as a solution and increased viscosity in gel form at ocular temperature. The optimized formulations, P7 (21% P407/4% P188), P8 (21% P407/5% P188) and F5 (1.5% chitosan and 45% ß-glycerophosphate) exhibited ideal ocular pH (7.27-7.46), phase transition at ocular temperature (33-37°C) and prolonged RA release up to 12 h. Formulation F5 showed an inhibition zone of 4.3 mm against M. luteus. Conclusion: Among all, formulation F5 alone exhibited modest antimicrobial activity against M. luteus. OS extracts at 5% and 10% were most active against tested bacteria however, loading them into in situ gels resulted in sedimentation. Hence, isolation of RA from OS extract is suggested before loading into formulations for a better antimicrobial activity.

Alleviation of Cadmium Stress by Silicon Supplementation in Peas by the Modulation of Morpho-Physio-Biochemical Variables and Health Risk Assessment.

Agricultural soil quality degradation by potentially toxic elements, specifically cadmium (Cd), poses a significant threat to plant growth and the health of humans. However, the supplementation of various salts of silicon (Si) to mitigate the adverse effect of Cd on the productivity of peas (Pisum sativum L.) is less known. Therefore, the present investigation was designed to evaluate the exogenous application at various levels (0, 0.50, 1.00 and 1.50 mM) of silicate compounds (sodium and potassium silicates) on pea growth, gaseous exchange, antioxidant enzyme activities and the potential health risk of Cd stress (20 mg kg-1 of soil) using CdCl2. The findings of the study showed that Cd stress significantly reduced growth, the fresh and dry biomass of roots and shoots and chlorophyll content. In addition, electrolyte leakage, antioxidant enzymes and the content of Cd in plant tissues were enhanced in Cd-induced stressed plants. An application of Si enhanced the development of stressed plants by modulating the growth of fresh and dry biomass, improving the chlorophyll contents and decreasing leakage from the plasma membrane. Furthermore, Si addition performed a vital function in relieving the effects of Cd stress by stimulating antioxidant potential. Hence, a significant level of metal protection was achieved by 1.00 mM of potassium silicate application under the Cd levels related to stress conditions, pointing to the fact that the Si concentration required for plant growth under Cd stress surpassed that which was required for general growth, enzymatic antioxidants regulation and limiting toxic metal uptake in plant tissues under normal conditions. The findings of this research work provide a feasible approach to reduce Cd toxicity in peas and to manage the entry and accumulation of Cd in food crops.

Molecular and Morphological Characterization of Eimeria crandallis Isolated from Deer (Cervidae) in Different Captive Animals.

Coccidiosis is a protozoan disease that is characterized by diffuse diarrhea, dehydration, emaciation accompanied by moderate morbidity and mild mortality in animals and birds. The current study targeted the molecular characterization of Eimeria isolates in captive deer from different localities in Lahore. The host species was the Cervidae family, such as Hog deer (Axis porcinus) and Punjab urial (Ovis aries vignei). The Eimeria crandallis was isolated from zoo animals. The DNA was extracted from oocysts and amplified by using reported oligonucleotide primers that exhibited the 809 bp product. These were analyzed by using the small subunit 18S rRNA gene-based evolutionary relationship with 36 other Eimeria species reported in caprine, cervinae, bovines, avians, and rodents. Light microscopic examination exhibited 3.29% (7/213) Eimeria-positive fecal samples with morphological features, including sub-spherical forms, the presence of micropyle with polar cap, and oocysts diameters (µm) ranging from 24.32 ± 1.61 to 18.94 ± 1.51. The phylogenetic tree constitutes four distinct clusters with relatively higher values. The evolutionary network showed that sequences were clustered in the monophyletic group of Eimeria species reported in caprine and cervinae. The nucleotide and amino acid sequence similarity matrix analysis exhibited 99.5-99.9% identity of the study isolates with Eimeria crandallis (AF336339). This study provides relevant baseline data to develop strategic control measures for coccidiosis in zoo animals. However, further investigations are required to place the hog deer and Punjab urial-derived E. crandallis into the caprine-originated cluster.

Propagation of Babesia bigemina in Rabbit Model and Evaluation of Its Attenuation in Cross-Bred Calves.

Bovine babesiosis (BB) is a vector-borne disease (VBD) that affects cattle in tropical and subtropical areas, caused by the haemoprotozoa Babesia bovis and Babesia bigemina. It is transmitted by tick bites belonging to the genus Rhipicephalus and is clinically characterized by high fever, depression, anorexia, decreased milk and meat production, haemoglobinemia, haemoglobinuria, jaundice, and pregnancy loss. In this study, the propagation of B. bigemina was evaluated by intraperitoneally inoculating 3 × 106 red blood cells infected with B. bigemina into rabbits. The study showed that variations in rabbit body temperatures are related to induced bovine babesiosis. A significant increase in temperature (39.20 ± 0.23 °C) was observed from day 4 onwards, with the maximum temperature (40.80 ± 1.01 °C) on day 9 post-inoculation. This study included susceptible cross-bred calves for in vivo attenuation, and they were compared with an infected group. The calves in the infected group showed a significant increase in temperature (38.79 ± 0.03 °C) from day 3 onwards and a maximum temperature (41.3 ± 0.17 °C) on day 11. Inoculated calves showed a gradual rise in temperature post-inoculation, but the difference was not significant. Inoculated calves did not show parasitaemia, whereas 32% of infected calves displayed parasitaemia. As compared to inoculated calves post-inoculation, packed cell volume (PCV) decreased (16.36 ± 1.30) for infected calves. However, there were statistically significant differences (p ≤ 0.05) in temperatures, parasitaemia, and PCV in both inoculated and infected calves. The current study aimed to attenuate B. bigemina in rabbit models and evaluate the pathogenic potential of this organism in naive calves. In conclusion, B. bigemina proliferation was attenuated in rabbits. The rabbit model can be used to study B. bigemina in vivo in order to reduce its pathogenicity.

Antibacterial, Anticandidal, and Antibiofilm Potential of Fenchone: In Vitro, Molecular Docking and In Silico/ADMET Study.

The aim of the present study is to investigate the effective antimicrobial and antibiofilm properties of fenchone, a biologically active bicyclic monoterpene, against infections caused by bacteria and Candida spp. The interactions between fenchone and three distinct proteins from Escherichia coli (ß-ketoacyl acyl carrier protein synthase), Candida albicans (1, 3-ß−D-glucan synthase), and Pseudomonas aeruginosa (Anthranilate-CoA ligase) were predicted using molecular docking and in silico/ADMET methods. Further, to validate the in-silico prediction, the antibacterial and antifungal potential of fenchone was evaluated against E. coli, P. aeruginosa, and C. albicans by determining minimum inhibitory concentration (MIC), minimum bacterial concentration (MBC), and minimum fungicidal concentration (MFC). The lowest MIC/MBC values of fenchone against E. coli and P. aeruginosa obtained was 8.3 ± 3.6/25 ± 0.0 and 266.6 ± 115.4/533.3 ± 230.9 mg/mL, respectively, whereas the MIC/MFC value for C. albicans was found to be 41.6 ± 14.4/83.3 ± 28.8 mg/mL. It was observed that fenchone has a significant effect on antimicrobial activity (p < 0.05). Our findings demonstrated that fenchone at 1 mg/mL significantly reduced the production of biofilm (p < 0.001) in E. coli, P. aeruginosa, and C. albicans by 70.03, 64.72, and 61.71%, respectively, in a dose-dependent manner when compared to control. Based on these results, it has been suggested that the essential oil from plants can be a great source of pharmaceutical ingredients for developing new antimicrobial drugs.

Nephroprotective effect of polyphenol-rich extract of Costus spicatus in cisplatin-induced nephrotoxicity in Wistar albino rats.

Costus spicatus (spiked spiralflag ginger) is traditionally utilised for its advantages such as antidiabetic, antihyperlipidemic, diuretic, antimicrobial, and anticancer properties. However, there is no scientific evidence on the nephroprotective potential of this plant. Thus, this study tested the nephroprotective effect of the polyphenol-rich extract of Costus spicatus leaves (PCSL) using preclinical models, including the HeK cell line and Wistar albino rats against cisplatin-induced toxicity. It also determined the polyphenolic compounds using high-performance thin-layer chromatography (HPTLC). PCSL showed significant (p < 0.05) nephroprotective potential against cisplatin-induced nephrotoxicity in HeK cells. Moreover, in vivo studies revealed significant (p < 0.05) amelioration in serum biochemical markers and antioxidant enzymes against cisplatin-induced nephrotoxicity. PCSL significantly inhibited the level of inflammatory cytokines such as TNF-α, IL-6, and IL-1ß. Moreover, PCSL restored the damage of the kidney tissues and ameliorated interstitial haemorrhage, congestion in capillaries, inflammatory cell infiltration, vacuolated cytoplasm, and tubular epithelial injury with widened Bowman's space. In addition, HPTLC analysis revealed that PCSL comprised polyphenolic compounds such as caffeic acid, quercetin, and ferulic acid. In conclusion, PCSL exerted nephroprotective potential by modulating the expression of inflammation, oxidative stress, and histological architecture of kidney tissues. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03233-z.

Silicon fertilization counteracts salinity-induced damages associated with changes in physio-biochemical modulations in spinach.

Plant growth and productivity are limited by the severe impact of salt stress on the fundamental physiological processes. Silicon (Si) supplementation is one of the promising techniques to improve the resilience of plants under salt stress. This study deals with the response of exogenous Si applications (0, 2, 4, and 6 mM) on growth, gaseous exchange, ion homeostasis and antioxidant enzyme activities in spinach grown under saline conditions (150 mM NaCl). Salinity stress markedly reduced the growth, physiological, biochemical, water availability, photosynthesis, enzymatic antioxidants, and ionic status in spinach leaves. Salt stress significantly enhanced leaf Na+ contents in spinach plants. Supplementary foliar application of Si (4 mM) alleviated salt toxicity, by modulating the physiological and photosynthetic attributes and decreasing electrolyte leakage, and activities of SOD, POD and CAT. Moreover, Si-induced mitigation of salt stress was due to the depreciation in Na+/K+ ratio, Na+ ion uptake at the surface of spinach roots, and translocation in plant tissues, thereby reducing the Na+ ion accumulation. Foliar applied Si (4 mM) ameliorates ionic toxicity by decreasing Na+ uptake. Overall, the results illustrate that foliar applied Si induced resistance against salinity stress in spinach by regulating the physiology, antioxidant metabolism, and ionic homeostasis. We advocate that exogenous Si supplementation is a practical approach that will allow spinach plants to recover from salt toxicity.

Molecular Characterization of Toxoplasma gondii in Cats and Its Zoonotic Potential for Public Health Significance.

Toxoplasmosis is a globally distributed disease of warm-blooded animals. It is caused by the opportunistic parasite Toxoplasma gondii (T. gondii). One-third of the global human population is believed to be infected with T. gondii. Cats serve as final host of T. gondii and are the main source of contamination of soil and water. This study aimed to detect genotypes of T. gondii in cats. Fecal samples (n = 400) were collected from districts of South Punjab (Khanewal and Sahiwal), and were processed by polymerase chain reaction (PCR) followed by sequencing and phylogenetic analysis. The obtained oligonucleotide sequences (T. gondii) were submitted to the GenBank database, and the evolutionary tree was constructed using MEGA-X software. Seven fecal samples (3.5%) from cats were positive. Five out of thirteen fecal samples (38.46%) found to be positive for T. gondii with microscopy were confirmed by PCR. After phylogenetic analysis with 3 clonal types and atypical strains, isolates of T. gondii in current study were more closely linked to a typical strain (AF249696). Besides genotyping from cats, seroprevalence from humans and ruminants is still considered to be the best and easiest way to identify the Toxoplasma. Blood samples were collected from sheep and goats (n = 2000 each), and human blood samples (n = 400) were collected from the same vicinity. Seroprevalence was determined using a commercial enzyme-linked immunosorbent assay (ELISA) kit. In Khanewal, the blood samples of 292 goats (29.2%) and 265 sheep (26.5%), and 6 fecal samples from cats (3%) were positive. Out of 200 human blood samples, 52 were positive, with a seroprevalence of 26%. In the Sahiwal district, the blood samples from 49 humans, 235 sheep and 348 goats were positive, with seroprevalence of 24.5%, 23.5% and 34.8%, respectively. The present study revealed the current circulating genotype of T. gondii from cats in the districts Khanewal and Sahiwal and the seroprevalence of the organism in small ruminants and humans living in the same vicinity. Further genotype analyses of the organism from ruminants and humans are needed.

A novel metabarcoded deep amplicon sequencing tool for disease surveillance and determining the species composition of Trypanosoma in cattle and other farm animals.

The World Health Organization (WHO) and the Food and Agriculture Organization (FAO) have developed strategies to control trypanosomiasis in humans and livestock in endemic areas. These require a better understanding of the distribution of different Trypanosoma species and improved predictions of where they might appear in the future, based on accurate diagnosis and robust surveillance systems. Here, we describe a metabarcoding deep amplicon sequencing method to identify and determine the Trypanosoma species in co-infecting communities. First, four morphological verified Trypanosoma species (T. brucei, T. congolense, T. vivax and T. theileri) were used to prepare test DNA pools derived from different numbers of parasites to evaluate the method's detection threshold for each of the four species and to assess the accuracy of their proportional quantification. Having demonstrated the accurate determination of species composition in Trypanosoma communities, the method was applied to determine its detection threshold using blood samples collected from cattle with confirmed Trypanosoma infections based on a PCR assay. Each sample showed a different Trypanosoma species composition based on the proportion of MiSeq reads. Finally, we applied the assay to field samples to develop new insight into the species composition of Trypanosoma communities in cattle, camels, buffalo, horses, sheep, and goat in endemically infected regions of Pakistan. We confirmed that Trypanosoma evansi is the major species in Pakistan and for the first time showed the presence of Trypanosoma theileri. The metabarcoding deep amplicon sequencing method and bioinformatics pathway have several potential applications in animal and human research, including evaluation of drug treatment responses, understanding of the emergence and spread of drug resistance, and description of species interactions during co-infections and determination of host and geographic distribution of trypanosomiasis in humans and livestock.

Molecular Evidence and Hematological Profile of Bovines Naturally Infected with Ehrlichiosis in Southern Punjab, Pakistan.

PURPOSE: Ehrlichiosis is one of the tick-borne diseases, which is not only neglected in Pakistan but only a few reports have been documented throughout the globe. The purpose of this study is to highlight and report the neglected pathogen from bovines in Pakistan. METHODS: In this study, the pathogen was detected initially based on microscopy, followed by the molecular confirmation and phylogenetic analysis of the pathogen from bovines from south Punjab, Pakistan. The hematological parameters were also assessed in Ehrlichia positive and negative animals. The information of different disease determinants was analyzed by a logistic regression model on SPSS. RESULTS: The study has reported an 11.98% (23/192) prevalence of Ehrlichiosis from bovines. The prevalence was slightly more in cattle (13.5%) as compared to the buffaloes (10.4%). The previous tick's history and tick control methods were proved to be the key factors with the occurrence of the disease. The isolates from Pakistan Ehrlichia spp. Pakistan/31, 36, and 8 clustered with the isolates from the USA, Nicaraguan, France, South Africa, and Uganda. Platelet count, hemoglobin level, and hematocrit were found significantly decreased in Ehrlichia affected animals when compared to the healthy bovines. CONCLUSION: This is the first report of ehrlichiosis from bovines in Pakistan and will provide the roadmap for future research.

Bioresource Nutrient Recycling in the Rice-Wheat Cropping System: Cornerstone of Organic Agriculture.

This study evaluated the impact of conventional practices (fertilizer alone) and diverse farming approaches (such as green manuring, farmyard manure application, rice-residue incorporation, residue mulching, residue removal and residue burning) on soil attributes. A total of thirty-five farm sites were selected, with five sites (replications) for each farming approach system, which were used over the past three years in the study farms. Characterization of rice residues of all cultivars, green manure crop (sesbenia: Sesbania sesban) and decomposed farmyard manure samples showed differential behaviours for macronutrients and micronutrients. Continuous application of inorganic fertilizers significantly influenced soil attributes, especially electrical conductivity, nutrient contents, bacterial and fungal population and soil enzymatic attributes. The crop residue treatments favourably influenced the soil parameters over the control. Crop residue incorporation or burning significantly increased soil available potassium, microbial biomass, enzymatic activities and organic carbon when compared with applications of chemical fertilizer alone, while total nitrogen content was increased by residue incorporation. However, green manuring and farmyard manure applications showed inferior responses compared with residue management treatment. It is therefore recommended that bioresources should be managed properly to warrant improvements in soil properties, nutrient recycling and the sustainability for crop productivity, in order to achieve sustainable development goals for climate action.

Silicon Fertilization: A Step towards Cadmium-Free Fragrant Rice.

Soil contamination with toxic cadmium (Cd) is becoming a serious global problem and poses a key hazard to environments and the health of human beings worldwide. The present study investigated the effects of foliar applications of three forms of silicate chemicals (calcium silicate, sodium silicate, and potassium silicate) at four rates (0.25%, 0.5%, 0.75%, and 1.0%) at tillering stage on rice growth and the accumulation of Cd under Cd stress (30 mg kg-1). The results showed that Cd stress reduced the yield-related traits and enlarged Cd contents in different rice organs. The leaf gas exchange attributes and yield traits were enhanced, and the Cd accumulation and bioaccumulation factor in rice organs were reduced, especially in grains, through silicon application. In shoots, roots, and grains, foliar spray of Si reduced Cd contents by 40.3%, 50.7%, and 47.9%, respectively. The effectiveness of silicate compounds in reducing Cd toxicity varied with the kind of chemicals and doses of foliar applications. Foliar application of potassium silicate, at a rate of 0.5%, at tillering stage, showed the best effectiveness in improving grain yield, while mitigating Cd accumulation in rice grains. The outcome of this study provides a promising practicable approach in alleviating Cd toxicity in rice and preventing the entrance of Cd into the food chain.

High-throughput sequencing of Fasciola spp. shows co-infection and intermediate forms in Balochistan, but only Fasciola gigantica in the Punjab province of Pakistan.

Fasciola gigantica and Fasciola hepatica are digenetic trematodes causing fasciolosis in ruminants. The host and geographical distribution of both Fasciola species are influenced by environmental and climatic conditions favouring survival and development of free-living stages and intermediate hosts, and livestock management practices. The aim of the present study was to describe the host distribution of the two Fasciola species in buffalo, cattle, goats, and sheep in the Balochistan and Punjab provinces of Pakistan. 359 flukes were collected from a total of 32 livers from the four livestock species. Deep amplicon sequencing of the internal transcribed spacer region 2 of ribosomal DNA (rDNA ITS-2) and mitochondrial nicotinamide adenine dinucleotide dehydrogenase 1 (mtDNA ND-1) loci confirmed co-infection of F. hepatica and F. gigantica in Balochistan and single species F. gigantica infection in Punjab. In Balochistan, co-infections and hybrids of both Fasciola species were identified in cattle, with more F. hepatica detected than F. gigantica. However, F. hepatica was the only species identified in goats, and F. gigantica was the only species identified in buffalo. In Punjab, all flukes were confirmed as F. gigantica in each of the four livestock species. Overall, the results indicate differences in the host and geographical distribution of F. gigantica and F. hepatica, and provide useful knowledge for the development of control strategies for livestock and humans.

An Overview of Phytochemical and Biological Activities: Ficus deltoidea Jack and Other Ficus spp.

Ficus deltoidea Jack (Moraceae) is a well-known medicinal plant used in customary medication among the Malay people to reduce and mend sicknesses such as ulcers, psoriasis, cytotoxicity, cardioprotective, inflammation, jaundice, vitiligo, hemorrhage, diabetes, convulsion, hepatitis, dysentery injuries, wounds, and stiffness. Ficus deltoidea contains a wide variety of bioactive compounds from different phytochemical groups such as alkaloids, phenols, flavonoids, saponins, sterols, terpenes, carbohydrates, and proteins. The genus Ficus has several hundreds of species, which shows excellent therapeutic effects and a wide variety of helpful properties for human welfare. Searching information was collected by using electronic databases including Web of Science, Science Direct, Springer, SciFinder, PubMed, Scopus, Medline, Embase, and Google Scholar. This review is, therefore, an effort to give a detailed survey of the literature on its pharmacognosy, phytochemistry, phytochemical, and pharmacological properties of Ficus and its important species. This summary could be beneficial for future research aiming to exploit the therapeutic potential of Ficus and its useful medicinal species.

In Vitro Cytotoxicity and Anti-inflammatory Cytokinine Activity Study of Three Isolated Novel Compounds of Prismatomeris glabra.

OBJECTIVES: The aim of the present study was to isolate and evaluate cytotoxicity and anti-inflammatory activities of new novel compounds isolated from Prismatomeris glabra. MATERIALS AND METHODS: Dried root of P. glabra was extracted under reflux with methyl alcohol, fractionated through the vacuum liquid chromatography technique, and evaporated and then purified the compounds using column chromatography and preparative thin-layer chromatography. THP-1 cells were treated with amentoflavone, 5,7,4'-hydroxyflavonoid, and stigmasterol with various concentrations (0-30 µg/mL) and then incubated with MTS reagent for 2h. Treatment was done for 24, 48, and 72h. Then, effects of these compounds were also tested on PGE2, TNF-α, and IL-6 expression in human THP-1-derived macrophage cells for 24h. RESULTS: Three new compounds such as amentoflavone, 5,7,4'-hydroxyflavonoid, and stigmasterol were isolated. After 24h of incubation, a significant decrease in cell viability was reported with IC50 values of amentoflavone, 5,7,4'- hydroxyflavonoid, and stigmasterol (21 µg/mL ≡ 38 M), (18 µg/mL ≡ 66 M) and (20 µg/mL ≡ 48.5 M), respectively. Whereas for 48 and 72h treatment showed a less decreased cell viability compared with 24h treatment. These compounds also showed a significant reduction in the production of TNF-α, IL-6, and PGE2 in a dose-dependent manner. CONCLUSIONS: The isolated new compounds showed significant cytotoxicity and anti-inflammatory effects.

Pharmacological Evaluation of Safoof-e-Pathar Phori- A Polyherbal Unani Formulation for Urolithiasis.

Safoof-e-Pathar phori (SPP) is an Unani poly-herbomineral formulation, which has for a long time been used as a medicine due to its antiurolithiatic activity, as per the Unani Pharmacopoeia. This powder formulation is prepared using six different plant/mineral constituents. In this study, we explored the antiurolithiatic and antioxidant potentials of SPP (at 700 and 1,000 mg/kg) in albino Wistar rats with urolithiasis induced by 0.75% ethylene glycol (EG) and 1% ammonium chloride (AC). Long-term oral toxicity studies were performed according to the Organization for Economic Co-operation and Development (OECD) guidelines for 90 days at an oral dose of 700 mg/kg of SPP. The EG urolithiatic toxicant group had significantly higher levels of urinary calcium, serum creatinine, blood urea, and tissue lipid peroxidation and significantly (p < 0.001 vs control) lower levels of urinary sodium and potassium than the normal control group. Histopathological examination revealed the presence of refractile crystals in the tubular epithelial cell and damage to proximal tubular epithelium in the toxicant group but not in the SPP treatment groups. Treatment of SPP at 700 and 1,000 mg/kg significantly (p < 0.001 vs toxicant) lowered urinary calcium, serum creatinine, blood urea, and lipid peroxidation in urolithiatic rats, 21 days after induction of urolithiasis compared to the toxicant group. A long-term oral toxicity study revealed the normal growth of animals without any significant change in hematological, hepatic, and renal parameters; there was no evidence of abnormal histology of the heart, kidney, liver, spleen, or stomach tissues. These results suggest the usefulness of SPP as an antiurolithiatic and an antioxidant agent, and long-term daily oral consumption of SPP was found to be safe in albino Wistar rats for up to 3 months. Thus, SPP may be safe for clinical use as an antiurolithiatic formulation.