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As cancer remains a significant global health challenge, there is an increasing need for novel therapeutic approaches. We investigated the antitumor potential of Juniperus communis berry essential oil on cervical cancer HeLa and colorectal HCT 116 cells. Cytotoxicity was evaluated through the MTT assay, revealing concentration-dependent reductions in cell viability. A clonogenic assay demonstrated long-term cytotoxic effects. Apoptosis markers were assessed via flow cytometric analysis and showed an induction of the intrinsic pathway in both cell lines, demonstrated by the elevated levels of cleaved caspase-3, Bax/Bcl-2 ratio, JC-10 monomer formation, and cytochrome C migration to the cytosol. The treatment inhibited cell-survival pathways in HCT 116 cells and arrested HeLa cells in the S phase. An extensive molecular docking screening provided insight into the binding affinity and interaction patterns of the essential oil components with NADH ubiquinone oxidoreductase and superoxide dismutase enzymes, further confirming the induction of the intrinsic pathway of apoptosis. The obtained in silico and in vitro results indicated the anticancer potential of J. communis berry essential oil as it interferes with cancer cell molecular mechanisms. Our findings highlight J. communis berry essential oil as a promising natural agent with anticancer potential.

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Of the six juniper species found in the Bulgarian flora, three of the species have controversial taxonomic positions. Juniperus pygmaea K. Koch and J. sibirica Burgsd. exhibit similar morphological characteristics to J. communis L. in terms of leaves and female cones (galbuli). This is one of the reasons why, in the recent taxonomic developments, J. pygmaea and J. sibirica were united in a common variety of J. communis, namely, J. communis var. saxatilis. However, such a grouping of species in the Flora of Bulgaria has not been adopted. This study aimed to evaluate the degrees of similarity or difference in the structure of the leaves, galbuli, seeds, and pollen of J. communis, J. sibirica, and J. pygmaea using the methods of comparative anatomy by light microscope (LM) and scanning electron microscopy (SEM) observations and complex morphological measurements. The working hypothesis of this study was that the three species would show a different degree of similarity with each other, which would clarify their taxonomic rank. The morphological parameters revealed differences between the length/width ratio of galbuli and seed length of the three species, while leaf characteristics (length and width) showed a stronger resemblance between J. sibirica and J. pygmaea. Furthermore, a greater distinction between the leaves and galbili of J. communis and J. sibirica was found. The SEM analyses showed variations in the seed shape and spermoderm among the three species. The shape of J. communis seeds was oval and elongated, while J. pygmaea seeds were pear-shaped, and J. sibirica seeds were triangular-rhombic. The length and height of striations were diverse on seed spermoderm in the three species. The epicuticular waxes of leaves, located on the tips of the anticlinal walls of the elongated epidermal cells in J. pygmaea and J. communis, were oval, while they formed raised comb-like crystals in J. sibirica. The morphological, anatomical, and SEM analysis affirmed the accepted taxonomic status of J. communis and J. sibirica as independent species within the Bulgarian flora. Based on most of the analyzed parameters, J. pygmaea exhibits significant similarity with J. sibirica. Additionally, the similar habitats of these two species support the determination of J. pygmaea as a variety or form of J. sibirica rather than J. communis (J. sibirica forma pygmaea).

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This study aims to elucidate the phytochemical diversity and biological activities of J. thurifera essential oil (JTEO) through a comparative analysis of samples from two distinct regions: Tensift-Al Haouz and Azilal, using both in vitro and in silico methods. Gas Chromatography-Mass Spectrometry (GC-MS) analysis revealed 21 components in the Tensift-Al Haouz JTEO (99.99% of the oil) and 23 components in the Azilal JTEO (99.58% of the oil), with oxygenated monoterpenes being the predominant compounds in both. The biological activities were assessed in vitro. Antioxidant properties, evaluated using DPPH, FRAP, and ABTS assays, showed significant activity in both oils. Antibacterial activity was tested against two strains of Gram-positive and two strains of Gram-negative bacteria, with both oils demonstrating notable bacterial growth inhibition. Enzymatic assays assessed the antidiabetic (α-amylase and α-glucosidase), dermo-protective (tyrosinase and elastase), and neuroprotective (AChE and BChE) activities. Both oils displayed substantial inhibitory effects across all tested activities, with variations attributed to their distinct chemical compositions. In silico analyses of six target enzymes confirmed significant binding affinities of the major compounds. Notably, 2,2'-Thiobis(6-tert-butyl-p-cresol) exhibited strong binding affinities with AChE, BChE, α-amylase, α-glucosidase, tyrosinase, and elastase, with binding energies ranging from -10.0 to -6.2 kcal/mol.

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During the screening of novel chemotherapeutic candidates from plants against adult T-cell leukaemia/lymphoma (ATL), we found that extracts of plants in the Solanaceae, Annonaceae, Apocynaceae, and Rutaceae families showed anti-proliferative activity in the MT-1 and MT-2 cell lines. We have isolated active compounds from these plants in the present research because Cupressaceae plants showed potent anti-proliferative activity in the cell lines. We attempted to isolate the active compounds from the leaves of Juniperus rigida. Using activity-guided fractionation, we isolated 22 compounds, including seven terpenoids, three aromatic compounds, two iridoids, five lignans, and five biflavonoids. The anti-proliferative activities of five diterpenoids were moderate, and those of two biflavonoids were stronger. A lignan (compound 13) showed the strongest activity. These compounds are promising candidates for the treatment of ATL.

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Plants interact with both antagonistic and mutualistic animals during reproduction, with the outcomes of these interactions significantly influencing plant reproductive success, population dynamics, and the evolution of plant traits. Here, we investigated the spatial and temporal variations in the interactions between Juniperus thurifera, its seed-dispersing birds, and three specific arthropod species that attack the fleshy cones during the predispersal period. We assessed how plant traits affect levels of cone damage by arthropods and seed dispersal by birds, the occurrence of competition among arthropod species, and the impact of seed predators on the activity of frugivores. Plant traits, cone damage by arthropods, and seed dispersal by birds showed spatiotemporal variability. Fluctuation in cone abundance was the leading factor determining damage by arthropods and bird dispersal with a secondary role of cone traits. Large crops satiated predispersal seed predators, although the amount of frugivory did not increase significantly, suggesting a potential satiation of bird dispersers. Crop size and cone traits at individual trees determined preferences by seed predator species and the foraging activity of bird dispersers. Competition among arthropods increased during years of low cone production, and seed predators sometimes negatively affected bird frugivory. High supra-annual variations in cone production appear to be a key evolutionary mechanism enhancing J. thurifera reproductive success. This strategy reduces the impact of specialized seed predators during years of high seed production, despite the potential drawback of satiating seed dispersers.

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Essential oils (EOs) are complex and susceptible to environmental conditions, they have a wide range of biological activities and are often used to differentiate between similar species. In this study, gas chromatography-mass spectrometry (GC-MS) coupled with chemometric analysis was applied to systematically analyse and evaluate EOs constituents and antioxidant activity of six Chinese Cupressaceae taxa (Platycladus orientalis Franco, P. orientalis Franco 'Sieboldii', P. orientalis Franco 'Aurea', Juniperus chinensis Roxb., J. chinensis Roxb. 'Kaizuca', and J. sabina L.) under identical conditions. The antioxidant activity of the EOs was evaluated using 2,2 -diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and ferric reducing power (FRAP), and the total phenolic content (TPC) of the EOs was determined by Folin-Ciocalteau reagent. In total, seventy individual constituents were identified with the main components being α-pinene, sabinene, D-limonene, bornyl acetate, δ-3-carene and ß-myrcene. Principal component analysis (PCA) and hierarchal cluster analysis (HCA) successfully discriminated the six taxa into three chemotypes and the unique chemotype revealed that J. chinensis 'Kaizuca' may be a species rather than a cultivar of J. chinensis. The results of OPLS-DA analysis showed that the three compounds screened, namely, α-pinene, sabinene, and δ-3-carene, can completely distinguish Platycladus spp. from Juniperus spp. The DPPH assay results ranged from 576.14 (J. chinensis 'Kaizuca') to 1146.12 (J. sabina) µmol eq Trolox/mL EO, while the ABTS values ranged from 1579.62 (P. orientalis 'Aurea') to 5071.82 (J. sabina) µmol eq Trolox/mL. In the FRAP assay, the values ranged from 1086.50 (J. chinensis 'Kaizuca') to 1191.18 (J. sabina) µmol eq Trolox/ml and the TPC of the EOs studied ranged from 15.17 (J. chinensis 'Kaizuca') to 39.37 (J. sabina) mg GAE/mL EO. The results consistently showed that J. sabina possessed the strongest antioxidant activity and can be preferentially used as a rich source of potentially natural antioxidants.

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Needle-leaved junipers (Juniperus sect. Juniperus, Cupressaceae) are coniferous trees and shrubs with red or blue fleshy cones. They are distributed across Asia, Macaronesia and the Mediterranean Basin, with one species (J. communis) having a circumboreal distribution. Here we aim to resolve the phylogeny of this clade to infer its intricate evolutionary history. To do so, we built a comprehensive, time-calibrated phylogeny using genotyping-by-sequencing (GBS) and combine it with species occurrence using phylogeographic tools. Our results provide solid phylogenetic resolution to propose a new taxonomic classification and a biogeographical history of the section. Specifically, we confirm the monophyly of two groups within J. sect. Juniperus: the Asian (blue-cone) species including the circumboreal J. communis, and the Mediterranean-Macaronesian (red-cone) species. In addition, we provide strong phylogenetic evidence for three distinct species (J. badia, J. conferta, J. lutchuensis) previously considered subspecies or varieties, as well as for the differentiation between the eastern and western Mediterranean lineages of J. macrocarpa. Our findings suggest that the Mediterranean basin was the primary center of diversification for Juniperus sect. Juniperus, followed by an East Asian-Tethyan disjunction resulting from uplifts of the Qinghai-Tibetan Plateau and climatic shifts. The colonization history of Macaronesia by red-cone junipers from the western Mediterranean appears to have taken place independently in two different geological periods: the Miocene (Azores) and the Pliocene (Madeira-Canary Islands). Overall, genomic data and phylogenetic analysis are key to consider a new taxonomic proposal and reconstruct the biogeographical history of the iconic needle-leaved junipers across the Paleartic.

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Habitat specialists have been largely overlooked in old-growth pinyon-juniper woodlands, despite specialists exhibiting heightened sensitivity to anthropogenic habitat loss. Furthermore, small mammal relationships within pinyon-juniper woodlands have most commonly been investigated via species abundance or habitat use, rather than habitat selection, thereby providing limited management metrics. We used the Oscura Mountains Colorado Chipmunk (Neotamias quadrivittatus oscuraensis) as a model organism to evaluate whether old-growth conditions drive resource selection by small mammals associated with pinyon-juniper woodlands. The goal of our study was to determine resources important to the chipmunk to inform management decisions. We evaluated microhabitat selection by testing a priori predictions based on natural history characteristics of the chipmunk and the woodlands. We grouped predictions into habitat characteristics affiliated with or not affiliated with old growth. We tested predictions under a multistage modeling framework using generalized linear mixed models with a binomial response variable of use versus availability. Probability of selection by chipmunks increased with increasing mean juniper diameter and increasing variation of pinyon diameter and decreased with increased distance to rocky escape terrain and increased mean percent grass cover. Our findings support the classification of the Oscura Mountains Colorado chipmunk as an old-growth pinyon-juniper specialist, as the chipmunk displayed disproportionate preference for old-growth microhabitat conditions. We recommend management policies that conserve old-growth multiage stands of pinyons and junipers. Old-growth conditions near outcroppings, escarpments, and large boulders are of particular conservation concern. Further, thinning resulting in increased grass cover may be detrimental to this old-growth pinyon-juniper specialist.

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Background and purpose: Cholestasis is caused by a malfunction of the biliary liver system. Oxidative stress plays an essential role in the progression of cholestasis. This study aimed to investigate the antioxidant and hepatoprotective effects of ethanolic extract of Juniperus excelsa M. Bieb (JE) fruits on hepatic impairment induced by bile duct ligation (BDL) in rats. Experimental approach: Forty male Wistar rats were randomly divided into 4 groups; sham control + vehicle (SC), BDL + vehicle (BDL), BDL + JE extract (BDL + JE), and SC + extract (SC + JE). One day after surgery, the animals were treated with vehicle or ethanolic extract of JE (500 mg/kg/day) for 7 days. Finally, the blood was taken for biochemical and oxidative stress analysis. Furthermore, the liver tissue of rats was removed for histological examination. Findings/Results: Treatment with the extract of JE decreased the ALP level, whereas it enhanced total protein content compared to the BDL group. Also, JE increased the activity of SOD and GPx, as well as FRAP content compared to the BDL group; while it did not significantly affect the levels of MDA and inflammation markers. However, JE could not improve BDL-induced histopathological alterations in hepatic tissue. Conclusion and implication: This study demonstrated that JE may be useful as an adjuvant therapy by attenuating ALP activity, increasing serum total protein and FRAP content, as well as improving the antioxidant enzymes activity of SOD and GPx. However, further research is warranted to explore the other underlying mechanisms of action.

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BACKGROUND: Asthma exacerbations are an important cause of emergency department visits but much remains unknown about the role of environmental triggers including viruses and allergenic pollen. A better understanding of spatio-temporal variation in exposure and risk posed by viruses and pollen types could help prioritize public health interventions. OBJECTIVE: Here we quantify the effects of regionally important Cupressaceae pollen, tree pollen, other pollen types, rhinovirus, seasonal coronavirus, respiratory syncytial virus, and influenza on asthma-related emergency department visits for people living near eight pollen monitoring stations in Texas. METHODS: We used age stratified Poisson regression analyses to quantify the effects of allergenic pollen and viruses on asthma-related emergency department visits. RESULTS: Young children (<5 years of age) had high asthma-related emergency department rates (24.1 visits/1,000,000 person-days), which were mainly attributed to viruses (51.2%). School-aged children also had high rates (20.7 visits/1,000,000 person-days), which were attributed to viruses (57.0%), Cupressaceae pollen (0.7%), and tree pollen (2.8%). Adults had lower rates (8.1 visits/1,000,000 person-days) which were attributed to viruses (25.4%), Cupressaceae pollen (0.8%), and tree pollen (2.3%). This risk was spread unevenly across space and time; for example, during peak Cuppressaceae season, this pollen accounted for 8.2% of adult emergency department visits near Austin where these plants are abundant, but 0.4% in cities like Houston where they are not; results for other age groups were similar. CONCLUSIONS: Although viruses are a major contributor to asthma-related emergency department visits, airborne pollen can explain a meaningful portion of visits during peak pollen season and this risk varies over both time and space because of differences in plant composition.

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Ethnobotanical studies revealed the experience and knowledge of people who learned the therapeutic virtues of plants through trials and errors and transferred their knowledge to the next generations. This study determined the ethnobotanical use of Juniperus drupacea (Andiz) in the Antalya province and the current and future potential distribution areas of J. drupacea in Türkiye during 2041-2060 and 2081-2100 according to the SSP2-4.5 and SSP5-8.5 scenarios and based on the IPSL-CM6A-LR climate change model. The very suitable areas encompassed 22379.7 km2. However, when the SSP2-4.5 scenario was considered, the areas most suitable for J. drupacea comprised 6215.892 km2 for 2041-2060 and 378.318 km2 for 2081-2100. Based on the SSP5-8.5 scenario, the area most suitable for J. drupacea was 979.082 km2 for 2041-2060. However, no suitable areas were identified with the SSP5-8.5 scenario for 2081-2100. Considering the models for the future estimated distribution areas of J. drupacea, serious contractions endangering the species are predicted in its distribution areas. Therefore, scientific research should focus on identifying J. drupacea populations and genotypes that demonstrate resilience to future drought conditions resulting from climate change. This endeavor is crucial as it holds significant ecological and economic values.

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The radial growth of trees in alpine timberline is particularly sensitive to climate change. We sampled and disposed tree-ring cores of three coniferous tree species including Juniperus saltuaria, Abies forrestii, and Larix potaninii at alpine timberline in Yading Nature Reserve. The standard tree-ring chronology was used to explore the response of radial growth of different timberline species to climate change. The results showed that radial growth of L. potaninii increased after 2000, while that of A. forrestii declined after 2002, and J. saltuaria showed a significant decreasing growth trend in the past 10 years. Such results indicated divergent growth responses to climate factors among the three tree species at alpine timberline. The radial growth of J. saltuaria was sensitive to temperature, and was positively correlated with the minimum temperature from previous October to current August, the mean tempera-ture from previous November to current April and from current July to October, but was negatively associated with the relative humidity from current July to October. The radial growth of A. forrestii showed negative correlation with mean temperature and the maximum temperature from May to June in the current year, while it exhibited positive association with the relative humidity and the Palmer drought severity index from May to June in the current year. L. potaninii radial growth was positively associated with mean temperature and the maximum temperature of November-December in the previous year, the maximum temperature of current March and mean temperature of current August. The temporal stability of climate-growth relationship varied among different timberline species. The positive correlation between radial growth of A. forrestii and J. saltuaria and temperature gradually decreased, while the posi-tive relationship of L. potaninii radial growth and temperature gradually increased. Under the background of climate warming, rapid rise in surface air temperatures may promote the radial growth of L. potaninii, while inhibit that of J. saltuaria and A. forrestii, which may change the position of regional timberline.

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The chemical composition of 33 oil samples isolated from the berries of individual trees of two subspecies of Juniperus oxycedrus L., collected in four locations in Tlemcen and Aïn Temouchent provinces, was investigated by GC-FID, GC/MS and 13CNMR. In total, 83 compounds were identified, accounting for 85.9 % to 98.6 % of the total composition. Yields ranged between 0.02 and 0.40 % (w/w). All compositions were submitted to statistical analysis. Combination of hierarchical clustering dendrogram and principal component analysis suggested the existence of two groups, the second group being sub-divided into two sub-groups. Groups and sub-groups were distinguished with respect to their contents of α-pinene, germacrene D, myrcene and, to a lesser extent, of manoyl oxide. The chemical composition of the oils belonging to subsp. oxycedrus was largely dominated by monoterpenes. α-Pinene (23.7-63.2 %) and germacrene D (4.5-25.4 %) were the major components, followed by myrcene (3.1-16.7 %). Conversely, samples oils of subsp. macrocarpa were largely dominated by germacrene D (4.4-30.0 %), followed by a lower content of α-pinene (10.0-24.3 %) and myrcene (1.0-4.3 %) and a significant content of manoyl oxide (4.3-9.9 %).

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Thalassochory, the dispersal of propagules through marine currents, is a key long-distance dispersal (LDD) mechanism with implications for global biogeography and particularly for island colonization. The propagules of coastal plant species are generally assumed to be better adapted for sea dispersal than those of inland plants, but this hypothesis remains largely untested. We conducted experiments on four genera (Juniperus, Daucus, Ferula, and Pancratium) and compared traits among nine species with different habitats and distributions. Our results showed that Juniperus spp. and P. maritimum have strong thalassochorous potential within the Mediterranean Basin. Interestingly, we did not find a clear association on the thalassochorous potential of coastal versus inland species within all the tested genera, apart from P. maritimum compared with the endemic inland P. illyricum. These findings suggest that thalassochory may be a more common dispersal mechanism than previously assumed. The apparently weak link of dispersal syndrome with species ecology broadens the possibility of dispersal by the sea also for inland plants, although considered to be poorly salt-tolerant. Moreover, our results reveal significant differences in sea dispersal between endemic and widespread species, but do not rule out an important role of thalassochory in shaping the distribution patterns of archipelago endemic flora. The presented method is largely replicable and could be used for further studies with a larger set of species to better delineate trends of sea dispersal syndrome among species with different ecology or dispersal traits.

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ETHNOPHARMACOLOGICAL RELEVANCE: Herbs of the genus Juniperus (family Cupressaceae) have been commonly used in ancestral folk medicine known as "Al'Araar" for treatment of rheumatism, diabetes, inflammation, pain, and fever. Bioassay-guided isolation of bioactives from medicinal plants is recognized as a potential approach for the discovery of novel drug candidates. In particular, non-addictive painkillers are of special interest among herbal phytochemicals. AIM OF THE STUDY: The current study aimed to assess the safety of J. thurifera, J. phoenicea, and J. oxycedrus aqueous extracts in oral treatments; validating the traditionally reported anti-inflammatory and analgesic effects. Further phytochemical investigations, especially for the most bioactive species, may lead to isolation of bioactive metabolites responsible for such bioactivities supported with in vitro enzyme inhibition assays. MATERIALS AND METHODS: Firstly, the acute toxicity study was investigated following the OECD Guidelines. Then, the antinociceptive, and anti-inflammatory bioactivities were evaluated based on chemical and mechanical trauma assays and investigated their underlying mechanisms. The most active J. thurifera n-butanol fraction was subjected to chromatographic studies for isolating the major anti-inflammatory metabolites. Moreover, several enzymatic inhibition assays (e.g., 5-lipoxygenase, protease, elastase, collagenase, and tyrosinase) were assessed for the crude extracts and isolated compounds. RESULTS: The results showed that acute oral administration of the extracts (300-500 mg/kg, p. o.) inhibited both mechanically and chemically triggered inflammatory edema in mice (up to 70% in case of J. thurifera) with a dose-dependent antinociceptive (tail flick) and anti-inflammatory pain (formalin assay) activities. This effect was partially mediated by naloxone inhibition of the opioid receptor (2 mg/kg, i. p.). In addition, 3-methoxy gallic acid (1), quercetin (2), kaempferol (3), and ellagic acid (4) were successfully identified being involved most likely in J. thurifera extract bioactivities. Nevertheless, quercetin was found to be the most potent against 5-LOX, tyrosinase, and protease with IC50 of 1.52 ± 0.01, 192.90 ± 6.20, and 399 ± 9.05 µM, respectively. CONCLUSION: J. thurifera extract with its major metabolites are prospective drug candidates for inflammatory pain supported with inhibition of inflammatory enzymes. Interestingly, antagonism of opioid and non-opioid receptors is potentially involved.

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Functional bowel disorders (FBD) have a major potential to degrade the standards of public life. Juniperus oxycedrus L. (J. oxycedrus) (Cupressaceae) has been described as a plant used in traditional medicine as an antidiarrheal medication. The present study is the first to obtain information on the antispasmodic and antidiarrheic effects of J. oxycedrus aqueous extract through in vitro and in vivo studies. An aqueous extract of J. oxycedrus (AEJO) was extracted by decoctioning air-dried aerial sections of the plant. Antispasmodic activity was tested in an isolated jejunum segment of rats exposed to cumulative doses of drogue extract. The antidiarrheic activity was tested using diarrhea caused by castor oil, a transit study of the small intestine, and castor oil-induced enteropooling assays in mice. In the jejunum of rats, the AEJO (0.1, 0.3 and 1 mg/ml) diminished the maximum tone induced by low K+ (25 mM), while it exhibited a weak inhibitory effect on high K+ (75 mM) with an IC50=0.49 ± 0.01 mg/ml and IC50=2.65 ± 0.16 mg/ml, respectively. In the contractions induced by CCh (10-6 M), AEJO diminished the maximum tone, similar to that induced by low K+ (25 mM). with an IC50=0.45 ± 0.02 mg/ml. The inhibitory effect of AEJO on low K+ induced contractions was significantly diminished in the presence of glibenclamide (GB) (0.3 µM) and 4-aminopyrimidine (4-AP) (100 µM), with IC50 values of 1.84 ± 0.09 mg/ml. and 1.63 ± 0.16 mg/ml, respectively). The demonstrated inhibitory effect was similar to that produced by a non-competitive antagonist acting on cholinergic receptors and calcium channels. In castor oil-induced diarrhea in mice, AEJO (100, 200, and 400 mg/kg) caused an extension of the latency time, a reduced defecation frequency, and a decrease in the amount of wet feces compared to the untreated group (distilled water). Moreover, it showed a significant anti-motility effect and reduced the amount of fluid accumulated in the intestinal lumen at all tested doses. These findings support the conventional use of Juniperus oxycedrus L. as a remedy for gastrointestinal diseases.

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Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of an essential oil obtained from the wood of Juniperus deppeana Steud. (cedarwood Texas oil), when used as a sensory additive for all animal species. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the essential oil under assessment is safe up to the maximum proposed use levels in complete feed of 15 mg/kg for veal calves (milk replacer), cattle for fattening, sheep, goats, horses, dogs, salmonids and ornamental fish. For the other species, the calculated safe concentrations in complete feed were 5 mg/kg for chickens for fattening, 8 mg/kg for laying hens, 7 mg/kg for turkeys for fattening, 10 mg/kg for piglets, 12 mg/kg for pigs for fattening, 14 mg/kg for sows and dairy cows, 8.5 mg/kg for rabbits and 4 mg/kg for cats. These conclusions were extrapolated to other physiologically related species. For any other species, the additive was considered safe at 4 mg/kg complete feed. The use of cedarwood Texas oil in water for drinking was considered safe provided that the total daily intake of the additive does not exceed the daily amount that is considered safe when consumed via feed. No concerns for consumers and the environment were identified following the use of the additive up to the maximum proposed use level in feed. The additive under assessment should be considered as irritant to skin and eyes, and as a skin and respiratory sensitiser. Since the individual components of cedarwood Texas oil are recognised to flavour food and their function in feed would be essentially the same as that in food, no further demonstration of efficacy was considered necessary.

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Background: This study presents the first faunistic record and DNA barcoding for some Diptera species recorded from the Juniperus forest ecosystem of Balochistan, Pakistan. DNA barcoding was used to explore species diversity of Dipterans and collections carried out using a Malaise trap between December 2018 to December 2019. This process involved sequencing the 658 bp Cytochrome Oxidase I (COI) gene. New information: Amongst the collected Diptera specimens, nine families were identified, representing 13 genera. These species include Atherigonasoccata (Rondani, 1871), Atherigonavaria (Schiner, 1868), Chironomusdorsalis (Meigen, 1818), Eupeodescorollae (Linnaeus, 1758), Eristalistenax (Linnaeus,1758), Goniaornata (Meigen, 1826), Luciliasericata (Meigen, 1826), Paragusquadrifasciatus (Linnaeus, 1758), Polleniarudis (Fabricius, 1794), Raviniapernix (Thompson, 1869), Sarcophagadux (Thompson, 1869), Trupaneaamoena (Schiner, 1868) and Wohlfahrtiabella (Linnaeus, 1758). The families Syrphidae and Sarcophagidae exhibited the highest representation, each comprising three genera and three species. They were followed by the family Muscidae, which had a single genus and two species. Anthomyiidae, Chironomidae, Calliphoridae, Polleniidae, Tachinidae and Tephritidae were represented by only one genus and one species. A nique Barcode Index Number (BIN) was allotted to Tachinidae (specie i.e Goniaornata). The results indicated that barcoding through cytochrome oxidase I is an effective approach for the accurate identification and genetic studies of Diptera species. This discovery highlights the significant diversity of this insect order in study region. Furthermore, a comprehensive list of other Diptera species remains elusive because of difficulties in distinguishing them, based on morphology and a lack of professional entomological knowledge.

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Juniperus, a genus of fragrant evergreen trees in the Cupressaceae family, encompasses up to 67 distinct species distributed globally. Among these, Juniperus procera, also known as the East African pencil cedar or African Juniper, stands out for its notable medicinal properties. Due to the well-recognized therapeutic benefits of Juniper species, assessing genetic diversity is essential for various breeding initiatives. Thus, in this work, six ISSR and six SCoT primers were utilized to evaluate the genetic diversity between 23 Juniper genotypes collected from different locations in Saudi Arabia. 29 out of 103 and 26 out of 105 amplified bands, respectively, were found to be polymorphic markers using the ISSR and SCoT studies. With the help of 120 genotype-specific markers, including 60 for ISSR and 60 for SCoT, several varieties of Juniper were discovered. In addition, the polymorphism information content (PIC) was computed to assess the effectiveness of the markers. The findings of this study highlight the importance of conserving the genetic diversity of Juniperus procera, as it holds immense potential for developing new medicinal products. Additionally, the results provide valuable insights into the genetic structure of Juniper populations in western Saudi Arabia, which can inform future conservation and management efforts. However, all of the techniques utilized to profile the genotypes of Juniper can be regarded as useful techniques for long-term fingerprinting and diagnostic markers.

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Purpose: Juniper (Juniperus procera) is a common forest tree species in Saudi Arabia. The decline in many populations of J. procera in Saudi Arabia is mainly due to seed dormancy and loss of natural regeneration. This study assessed the effects of chemical and hormonal treatments on seed germination and seedling growth in juniper plants. Methods: The seeds were subjected to either chemical scarification with 90% sulfuric acid and 20% acetic acid for 6 min or hormonal treatment by seed soaking in two concentrations (50 and 100 ppm) of three growth regulators, namely, indole acetic acid (IAA), gibberellins (GA3), and kinetin, for 72 h. A control group without any seed treatment was also prepared. The experiments were performed in an incubator maintained at room temperature and under a light and dark period of 12 h for 6 w. The germinated seeds for each treatment were counted and removed from the dishes. The selected germinated seeds from different treatments were planted in a greenhouse and irrigated with tap water for another 6 weeks. The hormone-treated seedlings were sprayed with their corresponding hormone concentrations 1 w after planting. Results: The highest percentage of seed germination was significantly recorded after seed soaking in 50 ppm GA3, whereas treatment with IAA (100 ppm) resulted in the best seedling growth. Seedlings treated with the three phytohormones showed a significant increase in photosynthetic pigments, total soluble sugars, proteins, percentage of oil, IAA, GA3, and kinetin contents of juniper seedlings compared with the control value, whereas abscisic acid content was decreased compared with chemical treatments. Conclusion: The investigated different treatments had an effective role in breaking seed dormancy and improving seedling growth of J. procera, which is facing a notable decline in its population worldwide. Moreover, such an effect was more pronounced in the three phytohormones that succeeded in breaking dormancy and growth of the Juniperus plant than in the other treatments.

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