Acute Effect of Caffeine on the Synthesis of Pro-Inflammatory Cytokines in the Hypothalamus and Choroid Plexus during Endotoxin-Induced Inflammation in a Female Sheep Model

This study was designed to determine the effect of acute caffeine (CAF) administration, which exerts a broad spectrum of anti-inflammatory activity, on the synthesis of pro-inflammatory cytokines and their receptors in the hypothalamus and choroid plexus (ChP) during acute inflammation caused by the injection of bacterial endotoxin—lipopolysaccharide (LPS). The experiment was performed on 24 female sheep randomly divided into four groups: control; LPS treated (iv.; 400 ng/kg of body mass (bm.)); CAF treated (iv.; 30 mg/kg of bm.); and LPS and CAF treated. The animals were euthanized 3 h after the treatment. It was found that acute administration of CAF suppressed the synthesis of interleukin (IL-1β) and tumor necrosis factor (TNF)α, but did not influence IL-6, in the hypothalamus during LPS-induced inflammation. The injection of CAF reduced the LPS-induced expression of TNF mRNA in the ChP. CAF lowered the gene expression of IL-6 cytokine family signal transducer (IL6ST) and TNF receptor superfamily member 1A (TNFRSF1) in the hypothalamus and IL-1 type II receptor (IL1R2) in the ChP. Our study on the sheep model suggests that CAF may attenuate the inflammatory response at the hypothalamic level and partly influence the inflammatory signal generated by the ChP cells. This suggests the potential of CAF to suppress neuroinflammatory processes induced by peripheral immune/inflammatory challenges.

Improvement in Tear Ferning Patterns of Sheep Tears After Addition of Various Electrolyte Solutions

Objective: This study aimed to improve the tear ferning (TF) patterns in the sheep tears after the addition of various electrolyte solutions in different proportions.Animal Studied: Sheep were located at a small farm in the outskirts of Riyadh, Saudi Arabia. The sheep had no ocular disorders or diseases, and none of the female sheep were pregnant.Methods: Tear samples (20 μl) were collected from the right eyes of seven healthy sheep (five female sheep and two male sheep; age 7–36 months with an average of 17.0 ± 10.3 months). A tear sample (1 μl) from each sheep was dried on a microscopic glass slide at 22°C and <40% humidity. The TF patterns were graded based on the five-point grading scale in 0.1 increments. Homogenous mixtures were prepared by mixing tears from each sheep (0.5 μl) with various electrolyte solutions in different proportions (1:1, 1:2, 1:4, 1:6, 1:8, and 1:10). A sample of each mixture (1 μl) was dried on a glass slide, and the TF patterns for each mixture were observed, recorded, graded, and compared with those of the corresponding pure sheep tears. In addition, each sheep tear sample (0.5 μl) was diluted with pure water (0.5 μl) and the TF images were recorded and graded to test the dilution effect.Results: General improvement was noted in TF grades after the addition of electrolyte solutions, ranging from 1.7–1.4 to 1.3–0.3 regardless of the ratio between the electrolyte solutions and sheep tears within the mixture. TF grades of sheep tear samples improved significantly after adding different volumes of calcium chloride solution. Similar improvements in TF grades were observed when magnesium chloride hexahydrate and sodium dihydrogen phosphate solutions were used as the electrolytes. Some improvements in the TF grades occurred with the addition of potassium chloride to sheep tear samples. There was little improvement in TF grades after the addition of sodium chloride solution.Conclusion: Tear ferning grades of sheep tear samples improved when mixed with a number of electrolyte solutions at different volumes, in particular with calcium chloride or magnesium chloride solutions. Some improvements in TF grades were seen with sodium dihydrogen phosphate or potassium chloride solution added as the electrolyte. Clearly, divalent electrolytes lead to a greater improvement in TF grades of sheep tear samples as compared with sodium dihydrogen phosphate or monovalent electrolytes.

Novel isolate of Cysticercus tenuicollis-Kalar isolate has been revealed in Kalar, Iraq

Introduction: Although Cysticercus tenuicollis is one of the most economic and veterinary important parasite in Iraq, scanty molecular characterization exists for this helminth. This study aimed to determine the prevalence and molecular description of C. tenuicollis isolates from sheep in Kalar district of Iraq. Methodology: A total of 2,906 slaughtered sheep were examined post-mortem. Up to 20 samples of C. tenuicollis was extracted and amplified using mitochondrial COX1 gene. Results: The overall prevalence rate was 6.88%, and female sheep recorded higher rate of infection (24.35%) than male (6.16%) with significant difference (p<0.05). The molecular results showed 14 haplotypes for COX1 gene and the pairwise nucleotide variation among them was ranged from 0.2 to 2.6%. Twelve out of fourteen haplotypes of C. tenuicollis involving one to three base mutations were discovered in Kalar, Iraq for the first time and this could be a unique mutation internationally and did not registered previously. Eleven newly recorded haplotypes involved only one single mutation and the remaining one involved three mutations. Phylogenetic interpretation showed that Cysticercus tenuicollis-Kalar isolate were clustered in one clade, and closely related to isolates discovered in Nigeria, China, Turkey, Poland, and Iran. Conclusions: This study provided a new record data on prevalence and discovered novel strains of C. tenuicollis in the study area for the first time named Cysticercus tenuicollis-Kalar isolate. Novel haplotypes might consider endemic genetic characterization of this metacestode. The present data may be useful to provide a good molecular background for future preventive and control programs.

Developmental Programming: Prenatal Testosterone Excess on Liver and Muscle Coding and Non-Coding RNA in Female Sheep

Prenatal testosterone (T)-treated female sheep manifest peripheral insulin resistance, ectopic lipid accumulation and insulin signaling disruption in liver and muscle. This study investigated transcriptional changes and transcriptome signature of prenatal T excess-induced hepatic and muscle-specific metabolic disruptions. Genome-wide coding and non-coding (nc) RNA expression in liver and muscle from 21-month-old prenatal T-treated (T propionate 100mg intramuscular twice weekly from days 30 to 90 of gestation; Term: 147 days) and control females were compared. Prenatal T (1) induced differential expression of mRNAs in liver (15 down, 17 up) and muscle (66 down, 176 up) (FDR&lt;0.05, absolute log2 fold change&gt;0.5); (2) downregulated mitochondrial pathway genes in liver and muscle; (3) downregulated hepatic lipid catabolism and PPAR signaling gene pathways; (4) modulated ncRNA metabolic processes gene pathway in muscle and (5) downregulated 5 uncharacterized long ncRNA (lncRNA) in the muscle but no ncRNA changes in the liver. Correlation analysis showed downregulation of lncRNAs LOC114112974 and LOC105607806 was associated with decreased TPK1, and LOC114113790 with increased ZNF470 expression. Orthogonal Projections to Latent Structures Discriminant Analysis identified mRNAs HADHA and SLC25A45, and miRNAs MIR154A, MIR25 and MIR487B in liver and ARIH1 and ITCH and miRNAs MIR369, MIR10A and MIR10B in muscle as potential biomarkers of prenatal T-excess. These findings suggest downregulation of mitochondria, lipid catabolism, and PPAR signaling genes in liver and dysregulation of mitochondrial and ncRNA gene pathways in muscle are contributors of lipotoxic and insulin resistant hepatic and muscle phenotype. Gestational T excess programming of metabolic dysfunctions involve tissue-specific ncRNA modulated transcriptional changes.

Kisspeptin, Neurokinin B, and Dynorphin Expression during Pubertal Development in Female Sheep

The neural mechanisms underlying increases in gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion that drive puberty onset are unknown. Neurons coexpressing kisspeptin, neurokinin B (NKB), and dynorphin, i.e., KNDy neurons, are important as kisspeptin and NKB are stimulatory, and dynorphin inhibitory, to GnRH secretion. Given this, we hypothesized that kisspeptin and NKB expression would increase, but that dynorphin expression would decrease, with puberty. We collected blood and hypothalamic tissue from ovariectomized lambs implanted with estradiol at five, six, seven, eight (puberty), and ten months of age. Mean LH values and LH pulse frequency were the lowest at five to seven months, intermediate at eight months, and highest at ten months. Kisspeptin and NKB immunopositive cell numbers did not change with age. Numbers of cells expressing mRNA for kisspeptin, NKB, or dynorphin were similar at five, eight, and ten months of age. Age did not affect mRNA expression per cell for kisspeptin or NKB, but dynorphin mRNA expression per cell was elevated at ten months versus five months. Thus, neither KNDy protein nor mRNA expression changed in a predictable manner during pubertal development. These data raise the possibility that KNDy neurons, while critical, may await other inputs for the initiation of puberty.

Impact of Extraction the Cause in Financial Transactions: A Comparative analytical Study: أثر تخريج المناط في المعاملات المالية: دراسة تحليلية مقارنة

The study aimed at showing practical impact which results from the process of extracting the cause in the matters of financial transactions. The researcher used for that the comparative analytical approach , where he turned to follow proofs of the very specific intentional financial transaction ; then, these proofs through abstract procedural and methodological tracks, to inspect the latent causes behind permission or prohibition of these transactions , in order these inferred doctrinal provisions to be adopted at the same course in relation to other transactions that take part with the same ones that their causes have been extracted. Undoubtedly, the type of transactions from which the researcher has taken the subject of his research that its cause is undiscussed, or what texts permit a decision without minimum sign to their cause; where the researcher studied the transaction of a man who purchases what he doesn't have, relying on proofs in this connection; then , he had inferred the cause of this transaction and extracted it , to arrive at the doctrinal impact arising from this extraction , i.e. giving the decision of this transaction itself to other transactions participating with it the cause as: sale of unawareness , sale before receiving , sale of non- existent , or there is not ability to deliver; and between unacceptability to include other transaction within the same arrangement , since the cause is different ; and an instance of that matter is giving back female sheep with a measure equals to the duration the purchaser makes benefit from the existence of this animal at his barn; and after extracting this cause of the matter , the researcher could establish a law of doctrinal impairments.

Epidemiological and Molecular Characterization of Echinococcus granulosus Isolated from Small Ruminants in Kashmir Valley, India

Background: Cystic Echinococcosis (CE) is an emergent or re-emergent zoonosis and remains a public health and economic problem all over the world. Methods: The present study was carried on the prevalence and genotypes of Echinococcus present in small ruminants in Kashmir valley. A total of 2100, sheep (2052) and goats (48), slaughtered or spontaneously dead, from various areas of Kashmir valley were screened for the presence of hydatidosis. In case of goat none of the cases were found positive for hydatidosis, whereas, all the positive cases (85) were recorded in sheep only. The overall prevalence of hydatidosis was 4.04%. The prevalence was higher in female sheep (5.46%) compared to males (2.83%). Season-wise highest prevalence was in summer (4.55%), followed by autumn (4.1%), spring (3.89%) and winter (2.5%).The liver was observed to be the most frequently infected organ with relative prevalence of 61.17% followed by lungs (38.82%).The rDNA-ITS1 fragment of positive samples was amplified with BD1 / 4S primers. Results: The length of amplified fragment for all isolated samples was 1000bps. The products obtained on PCR were digested with four restriction enzymes (Rsa 1, Alu 1, Msp 1 and Taq1). Rsa 1, Alu 1, Msp 1 yielded identical fragments, 300 and 700 bp in sheep. TaqI restriction enzyme had no effect on PCR product and after digestion; intact 1000bps fragment was seen. Conclusion: Phylogenetic analysis of ITS1 gene revealed that the common sheep strain (G1) is the predominant genotype in sheep in Kashmir valley.

Adipose Tissue- and Bone Marrow-Derived Mesenchymal Stem Cells from Sheep: Culture Characteristics

The current study demonstrates the culture characteristics of adipose tissue and bone marrow-derived mesenchymal stem cells (MSC). The study evaluates the effect of ambient temperature, physiological status of the donor and the tissue source on sheep (Ovis aries) mesenchymal stem cells. The tissue samples were harvested from full term pregnant female sheep (n = 9) and male sheep (n = 10). Adipose tissue was harvested from n = 9 sheep and bone marrow from n = 10 sheep. The samples (adipose tissue, n = 2; bone marrow, n = 3) transported at cold ambient temperature (<10 °C) failed to yield MSCs while those (n = 14) at higher (>20 °C) ambient temperature successfully yielded MSCs. Bone marrow mononuclear cell (MNC) fraction was higher than the adipose tissue-derived stromal vascular fraction (SVF), but the percent adherent cells (PAC) was higher in the later cell fraction. Adipose tissue-derived MSCs from the full term female sheep had a significantly (p < 0.05) higher proliferation potential as compared to those of the male sheep-derived MSCs. Female sheep MSCs also had rapid differentiation potential. The cryopreserved MSCs had morphological features comparable to that of the fresh cells. In conclusion, the tissue type and physiological status of donor animal may affect MSCs’ characteristics and should be taken into consideration while applying in clinical settings.

First molecular confirmation of Coenurus cerebralis in sheep and goats with neurological behaviors in Iraq

Background and Aim: Coenurus cerebralis is the intermediate stage of the canine cestode, which infect sheep and goats, resulting mainly in neurological signs and causing direct and indirect economic losses. This study aimed to demonstrate the existence of C. cerebralis and to elucidate the role of this parasite in inducing neurological behaviors in sheep and goats. Materials and Methods: On the basis of historical data on neurological signs, we subjected 76 animals (49 sheep and 27 goats) of different ages, sexes, and geographical areas for molecular examination of their blood samples using the polymerase chain reaction assay. Results: Of the 76 animals, 23.68% tested positive for C. cerebralis infection. We found significant increases in infection (p<0.05) in sheep (26.53%) more than in goats (18.52%). Circling movement was prevalent significantly among both C. cerebralis-positive sheep and goats. The Nineveh region had a significant (p<0.05) increase in positive sheep and goats, and the sheep of all study regions were infected significantly (p<0.05) more than were the goats. We found no significant (p>0.05) variation between sheep ages ≥1-3 years and ≥3 years; however, both groups had a significantly (p≤0.043) higher positivity rate than did sheep ages <1 year. The findings of sheep ages <1 year and ≥1-3 years were significantly higher than those of the goats, but not for goats ages ≥3 years. Female sheep and goats showed a significant increase in positivity versus that for the males. Conclusion: To the best of our knowledge, this study is the first report in Iraq targeting detection of C. cerebralis in sheep and goats with neurological behaviors; therefore, additional studies involving different animals in other regions using molecular techniques are needed.

The Effects of Vehicle Type, Transport Duration and Pre-Transport Feeding on the Welfare of Sheep Transported in Low Temperatures

Low temperatures can provide a risk to the welfare of sheep during transport because of increased ventilation chilling the sheep, and we examined the importance of three factors—covering the vehicle, duration of transport, and feeding prior to transport—on the welfare of sixty transported 4-month-old Dorper × Mongolian female sheep in a cold climate. Sheep in a covered vehicle had greater increases in head and ear temperatures than those in an open vehicle. Sheep transported for 2 h increased their leg temperatures, whereas those transported for 1 h had reduced leg temperatures. Increases in non-esterified fatty acids (NEFA) and lactate dehydrogenase (LDH) in the blood samples during the long transport suggested that sheep had more muscular and metabolic activity, compromising their well-being. Feeding prior to transport did not affect body temperatures, but those not fed prior to transport had reduced alanine transferase, HSP and cortisol in their blood, whereas those that were fed had reduced NEFAs, LDH and creatine kinase. Prior feeding had no effect on the sheep temperature indices over a two-hour transport period. Thus, the sheep most at risk of the adverse effects of cold temperatures were those transported in open vehicles, those transported for a longer time, and those not fed before transport.