Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

Application of touchdown enzyme time release (TETR)-PCR for diagnosis of Chlamydophila abortus infection

Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase chain reaction (PCR) assay as an improved test for sensitive and rapid diagnosis of abortion in small ruminants. Two hundred and fifty two placentae, liver or spleen tissue samples from aborting ewes and goats or aborted lambs and kids in which C. abortus infection was suspected were examined by TETR-PCR and the results were compared with cell culture. Sixty-five tissue samples were found to be TETR-PCR positive while only 56 samples were cell culture-positive. After resolution of discrepant samples with a confirmatory nested PCR assay, TETR-PCR had a sensitivity of 97% and a specificity of 99.5% while culture had a sensitivity of 84.8% and a specificity of 100%. The analytical sensitivity of the TETR-PCR assay was determined with DNA extracted from 4-fold serial dilution of C. abortus B577 culture and found to be 0.25 inclusion-forming unit per PCR. No reduction in the analytical sensitivity was noted when the assay was tested with mouse liver samples spiked with 4-fold serial dilution of C. abortus B577 culture. No target product was amplified when DNA from Chlamydophila pecorum was tested. TETR-PCR used in this study is a practical, rapid, sensitive and specific assay that could be used for the detection of C. abortus in infected tissue samples. We recommend the use of this assay as a supplemental diagnostic tool for detection of C. abortus in infected tissue samples.     

Evaluation of the Breeding Soundness of Male Camels (Camelus dromedarius) via Clinical Examination,Semen Analysis,Ultrasonography and Testicular Biopsy: A Summary of 80 Clinical Cases

Male camel infertility is a heterogeneous disorder. A variety of factors may adversely affect sperm production and function and impair fertility. This study was designed to evaluate the sensitivity and specificity of ultrasonography and testicular biopsy in the evaluation of the breeding soundness of male dromedaries compared with results obtained by clinical examination and semen analysis. Eighty‐four male dromedary camels (5–15 years old) were used in this study during the rutting season (November–May). Four sexually mature male camels were used as controls. These animals were apparently healthy and had histories of normal fertility. Eighty infertile male camels were subjected to an algorithmic approach based on information collected during careful examinations of the camels' breeding histories, clinical examinations, testicular evaluations, testicular ultrasonographies, the results of the semen analyses and testicular biopsies to diagnose the camels' infertilities. The differences in the semen parameters between the control and infertile male camels were highly significant (p < 0.01). Regarding the diagnoses of male camel infertility, the results of testicular ultrasonographies and biopsies were compared with those from the semen analyses, and the accuracies of these tests were 92.5% and 90%, respectively. Additionally, the results of the testicular ultrasonographies were matched with those of the testicular biopsies of the infertile animals, and this comparison resulted in 85% accuracy. Testicular biopsy is a promising method that, along with a carefully performed history, clinical examination, an appropriate testicular ultrasonography procedure and semen analysis, can afford veterinarians the opportunity for more precise diagnosis and treatment of many dromedary infertility disorders.     

Diagnosis of zinc deficiency in cotton

The diagnosis of Zn deficiency in cotton by means of total analysis or the appearance of deficiency symptoms is unsatisfactory. In order to obtain a better understanding of the development of Zn deficiency symptoms in relation to Zn content cotton plants were cultivated in waterculture under reproducible environmental conditions and Zn deficiency induced by different means. It was observed that deficiency symptoms developed equally, regardless of the inducing factors such as low Zn in the nutrient solution, high nutrient solution pH, high Ca and high Fe supply. High amounts of P in the nutrient solution did not induce Zn deficiency symptoms. Zn deficiency symptoms appeared first as interveinal chlorosis in the medium-aged leaves. The internodes remained short. Later red spots appeared on the leaf blades. The leaves suffering from deficiency were thicker because of enlarged palisade cells. The total Zn content of the leaves did not correlate with the symptoms. Different fractions of the Zn in the leaf (extractable with water, NaCl solution, hydrochloric acid) gave no better results. The analytically determined and calculated Zn fractions are discussed in relation to Zn deficiency inducing conditions.     

Addition of Leucaena-KX2 mulch in a shaded coffee agroforestry system increases both stable and labile soil C fractions

Agroforestry systems have the potential to increase sequestration of atmospheric carbon dioxide (CO₂) as soil organic carbon (SOC) because of the increased rates of organic matter addition and retention. However, few studies have characterized the relative stability of sequestered SOC in soil. We characterized SOC storage in aggregate size and chemical stability classes to estimate the relative stability of SOC pools after the addition of Leucaena-KX2 pruning residues (mulch) from 2006 to 2008 in a shaded coffee agroforestry system in Hawaii. Soil samples were separated by microaggregate isolation, density flotation and dispersion, and acid hydrolysis, resulting in five distinct fractions that differed in relative stability: coarse particulate organic matter (POM), fine POM, microaggregate-protected POM, silt + clay hydrolyzable soil organic matter (SOM), and silt + clay non-hydrolyzable SOM. With mulch addition, the fine POM fraction increased. There was also a shift in the proportion of SOC to more stable silt + clay fractions. In the absence of mulch there was no significant change in SOC fractions. Given that the turnover time of SOC in silt + clay fractions is on the order of decades to centuries, the potential benefits of active shade management and mulching compensate for the loss of C sequestration in tree biomass from pollarding.     

The flavonoid glycosides and procyanidin composition of Deglet Noor dates (Phoenix dactylifera)

The fruits of the date palm (Phoenix dactylifera) are consumed throughout the world and are an important part of the diet in the Middle East. Dates at the rutab and tamar maturity and ripening stages contain a wide array of phenolic antioxidants, but little is known about the composition of phenolic compounds in dates at the khalal stage of ripening. In the current study, the flavonoid glycoside and procyanidin compositions of dates of the cultivar Deglet Noor harvested at the khalal stage of maturity were characterized using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI/MS/MS). Procyanidin oligomers through decamers were identified in extracts of these dates. Higher molecular weight polymers, undecamers through heptadecamers, were also apparent from mass spectra. Thirteen flavonoid glycosides of luteolin, quercetin, and apigenin, 19 when considering isomeric forms, were also identified. Mass spectra indicate that both methylated and sulfated forms of luteolin and quercetin are present as mono-, di-, and triglycosylated conjugates whereas apigenin is present as only the diglycoside. LC-ESI/MS/MS spectra indicate that quercetin and luteolin formed primarily O-glycosidic linkages whereas apigenin is present as the C-glycoside.     

Acidulated activation of phosphate rock enhances release,lateral transport and uptake of phosphorus and trace metals upon direct-soil application

Phosphate rock (PR) was activated via acidulation with HCl, EDTA, and oxalic acid to enhance its reactivity. The release, lateral transport, and uptake of phosphorus (P) along with trace metals from pristine and activated PRs were investigated in a soil micro-block system over a period of 27 days, using wheat (Triticum aestivum L.) plants. Significantly (p < 0.05) higher amounts of available soil P, Fe, Mn, and Zn were released from all the PRs after application to soil within first 9 days of seedling transplantation, while the release of other trace metals (Cd, Co, Cr, Cu, Ni, and Pb) was minimal (<1.2 mg kg^?1). On cumulative basis, APR-O (oxalic acid activated PR) was the most efficient amendment releasing 164% more available P, followed by APR-E (EDTA activated PR) releasing 130% more available P, compared to the pristine PR. Similar results were also observed in the release of available Fe, Mn, Zn, and other trace metals. The highest diffusive mass fluxes for available P, Mn, Fe, and Zn in soil were observed after 3 days of seedling transplantation, which reduced subsequently. The uptake of P, Fe, Mn, and Zn by wheat plants was increased by 394%, 715%, 92%, and 91%, respectively, in APR-O application compared to the pristine PR, while it was increased by 280%, 188%, 16%, and 27%, respectively, in APR-E application compared to the pristine PR. Subsequently, APR-O and APR-E amendments resulted in enhanced shoot lengths, root lengths, shoot dry matter, and root dry matter contents of wheat plants. Hence, it was concluded that activation of PR with oxalic acid and EDTA prior to direct soil application may enhance the reactivity of PR and could serve as a cost-effect fertilization strategy for higher wheat crop production.     

Downregulation of male-specific cytochrome P450 by profenofos

The health hazards of individual organophosphorus insecticides have been characterized by their acute toxicity, mainly by investigating their cholinesterase inhibition. However, the chronic effects of most of these toxicants on the drug-metabolizing enzymes have not been investigated. Profenofos (O-4-bromo-2-chlorophenyl O-ethyl S-propyl phosphorothioate) is an organophosphorus pesticide widely used in cotton cultivation. In the present study, we investigated the effect of profenofos on male-specific cytochrome P450 (CYP) enzymes in adult Wistar rats. We orally administered 17.8 mg/kg body weight, twice weekly for 65 days. Profenofos downregulated levels of hepatic and testicular CYP2C11 and CYP3A2 mRNA and protein expression. Testicular aromatase (CYP19A) mRNA was decreased in the profenofos-treated rats compared to controls. Overall, the present study suggests that profenofos acts as an endocrine disruptor of male-specific CYP enzymes and affects testosterone concentration, which implicates its deleterious effects on animal or human males chronically exposed to organophosphorus pesticide.     

A comprehensive evaluation and first molecular report of Theileria ovis infection in small ruminants in Saudi Arabia

A total of 1000 clinically healthy small ruminants comprising 500 sheep and 500 goats from five districts within Riyadh Province in Saudi Arabia were investigated by routine Giemsa staining for hematozoan parasites. Out of these, 100 sheep and 95 goat samples were investigated by PCR using three pairs of hemoprotozoan-specific primers. Based on microscopic examination, 33.2% of sheep and 25.2% of goats were found infected with hemoprotozoan parasites, while PCR detected hematozoan infection in 46% of sheep and 33.7% of goats. Extensive molecular characterization of hematozoan infection using six pairs of species-specific primers revealed the dominance of Theileria ovis, rather than any other species, which is recorded for the first time in small ruminants in Saudi Arabia. Prevalence of T. ovis in sheep and goats was found to be the highest in Riyadh (32, 48%) followed by AL-Kharj (31, 35%), Ad-Dawadimi (31, 33%), AL-Majmaah (15, 27%), and Rumah (17, 23%). The highest parasite prevalence was recorded in the 3 years of age and >?4 years of age ruminants, while the lowest prevalence was recorded in <?1 year of age ruminants. No noticeable differences in parasite prevalence between male or female ruminants were recorded. Partial sequencing of 18S rRNA gene revealed the infection of the studied ruminants with four new isolates of T. ovis. Further characterization of the pathogenicity and the clinical effects of these T. ovis isolates in sheep and goats is highly needed. The current results can be helpful in protecting and improving livestock industry in the countries that depend on a high number of small ruminants.

     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.