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A comprehensive stress assessment is vital in understanding the impact of the pre-slaughter procedure on animal welfare. The transportation and handling process was commonly reported to cause stress in animals. This research utilises electroencephalography (EEG) as an alternative stress indicator to non-painful acute stress measurement. EEG has been proved to be instantaneous and sensitive with specific results. Therefore, this study was aimed to determine the stress level of goats subjected to two different transportation duration and the effect of lairage based on their EEG activities and blood parameters changes. Eighteen adult male goats were divided into two transportation stress groups based on the transport duration: the two-hour (TS2) and six-hour (TS6) groups. Then, each group was then again divided into three smaller groups according to the lairage duration, which was three-hour (L3), six-hour (L6), and overnight (L12) groups. Blood was sampled before transport, after transport, and during slaughter while EEG was recorded before transport, after transport, after lairage, and during slaughter. Results revealed that there was a significant decrease in beta wave activity compared to baseline in TS2 goats (P

Radiotherapy is one of the main options to cure and control breast cancer. The aim of this study was to investigate the sensitivity of two human breast cancer cell lines, MCF7 and MDA-MD-231, to radiation exposure at timepoints 4 h and 24 h after radiation. MCF7 and MDA-MD-231 were irradiated with different radiation doses using a Gilardoni CHF 320 G X-ray generator (Mandello del Lario, Italy) at 250 kVp, 15 mA [with half-value layer (HVL) = 1.6 mm copper]. The ApoTox-Glo triplex assay combines three assays used to assess viability, cytotoxicity, and apoptosis. The expression of γH2AX and BAX was analyzed by Western blotting. Viability and cytotoxicity did not change 4 h and 24 h after irradiation in either cell line, but we found a significant increase in the expression of cleaved caspase-3/7 at 24 h after irradiation with 8.5 Gy in MDA-MB231. The expression of γH2AX and BAX was low in MCF7, whereas the expression of γH2AX and BAX increased with radiation dose in a dose-dependent manner in MDA-MB231. The results show that the MCF7 cell line is more radioresistant than the MDA-MB 231 cell line at 4 h and 24 h after X-ray irradiation. In contrast, MDA-MB-231 cells were radiosensitive at a high radiation dose of 8.5 Gy at 24 h after irradiation. γH2AX and BAX indicated the radiosensitivity in both cell lines. These results open the possibility of using these cancer cell lines as models for testing new therapeutic strategies to improve radiation therapy.

As of today, no literature has been reported on the efficacy of stevia on lipid regulations conducted in vitro. Thus, the current study was focusing on the potential of Stevia rebaudiana bertoni as an anti-hypercholesterolemia substitute in limiting the de novo cholesterol synthesis in Hep-G2 cell line. The cytotoxicity and lipid internalization effects of stevia on Hep-G2 cells were assessed quantitatively and qualitatively in this study. As evaluated by MTT assay, commercialized stevia (0.5-20.0 mg/ml) and stevioside (1.0-10 µM) inhibited Hep-G2 cells viability in a dose-dependent manner for 24 hours. IC50 was detected at 8.68 mg/ml (commercialized stevia) and 10.91 µM (stevioside). From the assay, suitable concentrations were chosen to study the effect of stevia on cholesterol internalization in Hep-G2 cells supplemented with exogenous lipids. Cholesterol quantification assay revealed that high concentration commercialized stevia and stevioside promoted significant cholesterol internalized in Hep-G2 cells as compared to simvastatin. Finally, immunofluorescent microscopy assessment was done to qualitatively observe the formation of lipid droplets and low-density lipoprotein receptor in relation to total cholesterol extracted. The microphotographs of immunofluorescent microscopy were in parallel to results obtained from the cholesterol quantification assay which further revealed the effect of stevia as a potential anti-hypercholesterolemia agent.

Melastoma malabathricum is a well-known herb in Malaysia where it being used in various ways for treatment of different diseases and ailments including skin problems. The study aims to investigate acute and subacute dermal toxicity of ethanolic extract of M. malabathricum leaves following to a single or repeated doses exposure. A total of 30 female Sprague-Dawley rats were grouped into 5 groups (n = 6 per group) for both acute and subacute toxicity study. The duration for each study was determined at 14 days for acute toxicity and 28 days for subacute toxicity. The rats were topically applied with the plant extract at three different doses; 2.5%, 5.0% and 10.0% on the shaved area of dorsal skin. For acute toxicity study, rats in all three groups received single application of the extract on the first day of the experimental period, while rats in subacute toxicity study were topically applied with the extract once daily for 28 days. Throughout the respective 14-day and 28-day study periods, all rats were monitored for any changes in their physical appearance and behavioural patterns that might develop due to toxic effects of the plant. There were no mortality or abnormal physical appearance, and physiological and behavioural changes observed in all rats in both studies. Body weights, kidney and liver weights, and both haematology and serum biochemistry results showed no significant (p > 0.05) differences between all groups in both studies. All of the findings were supported by normal macroscopic and microscopic architectures of liver, kidneys and skin of all rats applied topically with the extract. This study suggests that topical application of M. malabathricum leaf ethanolic extract at 2.5%, 5% and 10% does not induce acute and subacute adverse effects on the skin or systemic toxic reactions in rats.

This study was aimed to provide the reference frame for the safe dose design of polyphenol-rich herbal mixture, which consist of lemon, apple cider, garlic, ginger and honey (PRM) for the future efficacy study. Prior to this, the nutritional composition was first conducted and the identification of metabolites that present in PRM was determined using proton nuclear magnetic resonance (1H-NMR) spectroscopy. The acute toxicity of the PRM was then evaluated in zebrafish embryo and Wistar rats following The Organisation for Economic Co-operation and Development (OECD) guidelines. The PRM nutritional composition and sugar profile showed it was high in carbohydrate, ash and protein and the main sugar is fructose. It also contains metabolites such as fructosefuranose, lactic acid, ascorbic acid, acetic acid, cycloalliin, pyruvate, 5- hydroxymethylfurane, α- and β-glucose. From the zebrafish embryo acute toxicity result, the lethal concentration, LC50 of PRM was at 487.50 μg/mL. Meanwhile, in Wistar rats’ model, no lethality was observed in the group treated with PRM at the end of the study (14 days). No changes were also observed from the behavioural and appetite as well as the biochemical parameters (AST, ALT, total cholesterol, triglyceride, LDL, HDL, total protein and creatinine) of the treated group. Therefore, the safe dose for PRM can be up to 2000 mg/kg b.w. in Wistar rats and below 487.50 μg/mL in zebrafish embryo model.

An herbal mixture composed of lemon, apple cider, garlic, ginger and honey as a polyphenol-rich mixture (PRM) has been reported to contain hypolipidemic activity on human subjects and hyperlipidemic rats. However, the therapeutic effects of PRM on metabolites are not clearly understood. Therefore, this study aimed to provide new information on the causal impact of PRM on the endogenous metabolites, pathways and serum biochemistry. Serum samples of hyperlipidemic rats treated with PRM were subjected to biochemistry (lipid and liver profile) and hydroxymethylglutaryl-CoA enzyme reductase (HMG-CoA reductase) analyses. In contrast, the urine samples were subjected to urine metabolomics using 1H NMR. The serum biochemistry revealed that PRM at 500 mg/kg (PRM-H) managed to lower the total cholesterol level and low-density lipoprotein (LDL-C) (p &lt, 0.05) and reduce the HMG-CoA reductase activity. The pathway analysis from urine metabolomics reveals that PRM-H altered 17 pathways, with the TCA cycle having the highest impact (0.26). Results also showed the relationship between the serum biochemistry of LDL-C and HMG-CoA reductase and urine metabolites (trimethylamine-N-oxide, dimethylglycine, allantoin and succinate). The study’s findings demonstrated the potential of PRM at 500 mg/kg as an anti-hyperlipidemic by altering the TCA cycle, inhibiting HMG-CoA reductase and lowering the LDL-C in high cholesterol rats.

Stevia rebaudiana Bertoni is a native plant to Paraguay. The extracts have been used as a famous sweetening agent, and the bioactive components derived from stevia possess a broad spectrum of therapeutical potential for various illnesses. Among its medicinal benefits are anti-hypertensive, anti-tumorigenic, anti-diabetic, and anti-hyperlipidemia. Statins (3-hydro-3-methylglutaryl-coenzyme A reductase inhibitor) are a class of drugs used to treat atherosclerosis. Statins are explicitly targeting the HMG-CoA reductase, an enzyme in the rate-limiting step of cholesterol biosynthesis. Despite being widely used in regulating plasma cholesterol levels, the adverse effects of the drug are a significant concern among clinicians and patients. Hence, steviol glycosides derived from stevia have been proposed as an alternative in replacing statins. Diterpene glycosides from stevia, such as stevioside and rebaudioside A have been evaluated for their efficacy in alleviating cholesterol levels. These glycosides are a potential candidate in treating and preventing atherosclerosis provoked by circulating lipid retention in the sub-endothelial lining of the artery. The present review is an effort to integrate the pathogenesis of atherosclerosis, involvement of lipid droplets biogenesis and its associated proteins in atherogenesis, current approaches to treat atherosclerosis, and pharmacological potential of stevia in treating the disease.

Animal models have been providing invaluable contributions to the better understanding of mechanisms of cancer (including leukaemias) development and effectiveness of most of the treatments. Chemical carcinogens are generally used to study the biology of cancers including leukaemias in many animal models, including rats and mice. The studies in most cases are aimed at the development and evaluation of cancer treatments and preventions. Some of the most common chemical carcinogens used in animal models for leukaemias include N-ethyl-N-nitrosourea (ENU), N-methyl-N-nitrosourea (MNU), dimethyl benz(a)anthracene (DMBA) and benzo(a)pyrene (BaP). This review provides highlights on different animal models of leukaemia induced by the chemical carcinogens mentioned earlier, at the same time discussing the contributions of these models to the leukaemia diagnosis in laboratory animal models for subsequent development of treatment.

The aim of this study was to measure the serum proinflammatory (IL-12, GM-CSF &amp, IFN-γ) to anti-inflammatory (IL-10, IL-4) cytokine ratio, oxidant (MDA) level and antioxidant enzyme (SOD, GPx) activities after blood parasite infections. The blood and serum samples were obtained from 130 cattle and screened for identity of the infecting blood parasites by conventional PCR. The following blood parasite species were detected: Candidatus Mycoplasma haemobos (70/130), Theileria orientalis (65/130), Theileria sinensis (32/130), Anaplasma marginale (49/130), Anaplasma platys (7/130), and Trypanosoma evansi (4/130). The GM-CSF/IL-10 ratio showed significantly higher values in all the symptomatic blood parasite infected cattle groups except for symptomatic A. platys infected cattle groups. Anti-inflammatory cytokine immune responses were notable findings in symptomatic and asymptomatic cattle infected with C. M. haemobos and T. orientalis characterized by low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 (p &lt, 0.05). Therefore, high serum GM-CSF:IL:10 in the symptomatic blood parasite infected cattle, low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 ratios in asymptomatic cattle, high MDA level, and increased antioxidant enzyme activities could be useful predictive tools for outcome of natural blood parasite infections in cattle.

The Thai Journal of Veterinary Medicine, 51, 1, 101

Plastic debris is widespread and ubiquitous in the marine environment and ingestion of plastic debris by marine organisms is well-documented. Viscera and gills of 110 individual marine fish from 11 commercial fish species collected from the marine fish market were examined for presence of plastic debris. Isolated particles were characterized by Raman spectroscopy, and elemental analysis was assessed using energy-dispersive X-ray spectroscopy (EDX). Nine (of 11) species contained plastic debris. Out of 56 isolated particles, 76.8% were plastic polymers, 5.4% were pigments, and 17.8% were unidentified. Extracted plastic particle sizes ranged from 200 to 34,900 μm (mean = 2600 μm ±7.0 SD). Hazardous material was undetected using inorganic elemental analysis of extracted plastic debris and pigment particles. The highest number of ingested microplastics was measured inEleutheronema tridactylumandClarias gariepinus, suggesting their potential as indicator species to monitor and study trends of ingested marine litter.

Background Brucellosis of goats is caused by Brucella melitensis. It is a re-emerging zoonotic disease in many countries due to transmission from domestic animals and wildlife such as ibex, deer and wild buffaloes. Objective To describe the pathological changes, identification and distribution of B. melitensis in foetuses of experimentally infected does. Methods Twelve female goats of approximately 90 days pregnant were divided into 4 groups. Group 1 was exposed intra-conjunctival to 100 µL of sterile PBS while goats of Groups 2, 3 and 4 were similarly exposed to 100 µL of an inoculum containing 109 CFU/mL of live B. melitensis. Goats of these groups were killed at 15, 30 and 60 days post-inoculation, respectively. Foetal fluid and tissues were collected for bacterial identification (using direct bacterial culture, PCR and immuno-peroxidase staining) and histopathological examination. Results Bilateral intra-conjunctival exposure of pregnant does resulted in in-utero infection of the foetuses. All full-term foetuses of group 4 were either aborted or stillborn, showing petechiations of the skin or absence of hair coat with subcutaneous oedema. The internal organs showed most severe lesions. Immune-peroxidase staining revealed antigen distribution in all organs that became most extensive in group 4. Brucella melitensis was successfully isolated from the stomach content, foetal fluid and various other organs. Conclusion Vertical transmission of caprine brucellosis was evident causing mild to moderate lesions in different organs. The samples of choice for isolation and identification of B. melitensis are stomach content as well as liver and spleen tissue.

Background Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos and Trypanosoma evansi, and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle. Methods Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, MPSP genes of T. orientalis and T. sinensis, MSP4 gene of A. marginale, 16SrRNA gene of Candidatus Mycoplasma haemobos, and RoTat1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia. Results A total of 44 (72.13%) cattle were infected with more than one blood pathogen. Theileria species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83–81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98–100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16SrRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of T. sinensis. The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the T. evansi and Anaplasma platys + Theileria sinensis double species co-infected cattle group. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p T. evansi infected cattle when compared to clinically healthy cattle. Conclusion We present the first evidence of Theileria sinensis-associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of A. platys, there is an urgent need for appropriate preventive and control measures against these blood pathogens.

Theileriosis is a blood piroplasmic disease that adversely affects the livestock industry, especially in tropical and sub-tropical countries. It is caused by haemoprotozoan of the Theileria genus, transmitted by hard ticks and which possesses a complex life cycle. The clinical course of the disease ranges from benign to lethal, but subclinical infections can occur depending on the infecting Theileria species. The main clinical and clinicopathological manifestations of acute disease include fever, lymphadenopathy, anorexia and severe loss of condition, conjunctivitis, and pale mucous membranes that are associated with Theileria-induced immune-mediated haemolytic anaemia and/or non-regenerative anaemia. Additionally, jaundice, increases in hepatic enzymes, and variable leukocyte count changes are seen. Theileria annulata and Theileria parva induce an incomplete transformation of lymphoid and myeloid cell lineages, and these cells possess certain phenotypes of cancer cells. Pathogenic genotypes of Theileria orientalis have been recently associated with severe production losses in Southeast Asia and some parts of Europe. The infection and treatment method (ITM) is currently used in the control and prevention of T. parva infection, and recombinant vaccines are still under evaluation. The use of gene gun immunization against T. parva infection has been recently evaluated. This review, therefore, provides an overview of the clinicopathological and immunopathological profiles of Theileria-infected cattle and focus on DNA vaccines consisting of plasmid DNA with genes of interest, molecular adjuvants, and chitosan as the most promising next-generation vaccine against bovine theileriosis.

Background: Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Trypanosoma evansi, Theileria, Anaplasma and Mycoplasma species, and studied their phylogenetic relationships, haemato-biochemical abnormalities and erythrocyte osmotic fragility (EOF) in Malaysian cattle. Methods: Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, 16SrRNA gene of Candidatus Mycoplasma haemobos, MPSP gene of T. orientalis and RoTaT1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) Kedah-Kelantan X Brahman cattle from Pahang, Malaysia. Haemato-biochemical analyses were performed using automated analysers while EOF was determined with the aid of saline solutions. Results: PCR amplification produced the expected fragment sizes for MPSP gene of T. orientalis, msp4 gene of Anaplasma marginale, 16S rRNA gene of C. M. haemobos, RoTaT1.2VSG gene of T. evansi. Nucleotide blast demonstrated that sequences of the PCR amplicons showed a high degree of molecular similarity in comparison with reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16S rRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys as additional species detected in these cattle. Theileria species was the most detected blood pathogen in the sampled cattle. The blood picture of all cattle group revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. Erythrocyte fragility values of all the cattle groups were above the reference range. Anaemia of the macrocytic normochromic type was observed in the Trypanosoma evansi; and Anaplasma platys + Theileria sinensis double species co-infected cattle. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (pTrypanosoma evansi infected cattle.Conclusion: We present the first report of anaemia associated with Theileria sinensis infection in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of Anaplasma platys, there is an urgent need for appropriate preventive and control measures, as Theileria species and A. platys are of great economic and zoonotic importance respectively.

Background: breast cancer is one of most common types of cancer and heterogeneous disease. Somatostatain receptors (SSTR1-5) are expressed in breast cancer cells. MCF7 and MDA-MB231 cell lines used to evaluate the expression of somatostatin receptors (SSTRs 1 - 5). Methods: The detection of mRNA expression levels of SSTRs in MCF7 and MDA-MB231 was performed using quantitative- polymerase chain reaction (qPCR). Results: All SSTRs 1 - 5 were expressed in both cells lines. The SSTR 1, 2, 3 and 4 mRNA levels were significantly higher in MDA-MB-231 in relation to MCF-7. The expression of SSTR 4 and 5 mRNA was highest in MDA-MB231 and MCF-7 cell lines respectively, however, SSTR3 mRNA was least expressed in both cell lines. Conclusion: All SSTRs were expressed in both MDA-MB231 and MCF7 cells. However, the levels of expression differ between both cell lines. Keywords: Breast cancer. Cell lines .Somatostatin receptors. mRNA expression

Chemical carcinogens are commonly used to investigate the biology and prognoses of various cancers. This study investigated the mechanism of leukaemogenic effects of n-ethyl-n-nitrosourea (ENU) in a mouse model. A total of 14 3-week-old male Institute of Cancer Research (ICR)-mice were used for the study. The mice were divided into groups A and B with seven mice each. Group A served as the control while group B received intraperitoneal (IP) injections of 80 mg/kg ENU twice with a one-week interval and were monitored monthly for 3 months for the development of leukaemia via blood smear examination. The mice were sacrificed humanely using a CO2 chamber. Blood, spleen, lymph nodes, liver, kidney and lung samples were collected for blood smear examination and histopathological evaluation. The expression of angiogenic protein (VEGF), and pro and anti-apoptotic proteins (BCL2 and BAX), was detected and quantified using Western blot technique. Leukaemia was confirmed by the presence of numerous blast cells in the peripheral blood smear in group B. Similarly, the VEGF and BCL2 proteins were significantly (p < 0.05) upregulated in group B compared to A. It is concluded that IP administration of 80 mg/kg ENU induced leukaemia in ICR-mice 12 weeks post administration through upregulation of angiogenic and anti-apoptotic proteins: VEGF and BCL2.

Hypercholesterolemia is a major risk factor for the initiation and development of nonalcoholic fatty liver disease and atherosclerosis. The present study evaluated the hypocholesterolemic effect of virgin avocado oil (VAO) using urinary metabolomic method. Male Sprague-Dawley rats were fed high-cholesterol diet for four weeks to induce hypercholesterolemia. After confirming the establishment of hypercholesterolemia model, the VAO (450 and 900 mg·kg-1 ·day-1 ) and simvastatin (10 mg·kg-1 ·day-1 ) were given orally while maintaining the high-cholesterol diet for another four weeks. Assessment of urinary metabolomics using NMR revealed that VAO treatment could partially recover the metabolism dysfunction induced by hypercholesterolemia mainly via lipid, energy, amino acid, and gut microbiota metabolism.

Avocado (Persiana americana Mill) belongs to the Lauraceae family. High level of lipids in the avocado pulp can be served as an important raw material for edible oil extraction. In the present study, the physicochemical properties of avocado oil extracted using subcritical CO2 extraction (SCO2) and ultrasound-assisted aqueous extraction (UAAE) were compared with the conventional solvent extraction. In comparison to solvent extraction, the oils extracted using SCO2 and UAAE were found to have higher iodine values, but lower slip melting points, free fatty acid contents and saponification values. Regardless of the extraction methods, the major fatty acids in avocado oils were oleic (40.73-42.72%) and palmitic (28.12-34.48%) acids whereas the major triacylglycerols in avocado oils were palmitoyl-dioleoyl-glycerol (POO; 22.48-23.01%) and palmitoyl-oleoyl-linoleoyl-glycerol (POL; 17.64-18.23%). SCO2 and UAAE are effective “solvent-free” methods to extract avocado oils and potentially other edible oils.

A nanocomposite, phytic acid-chitosan-magnetic iron oxide nanoparticles (IP6-CS-MNPs) has been used to treat colon cancer in vitro, previously. However, its potential toxicity in vivo has yet to be elucidated. Hence, the present study aimed to evaluate the acute effects of oral administration of IP6-CS-MNPs in mice. In this study, 1000 and 2000 mg/kg body weight (b.w) of IP6-CS-MNPs were orally administered to two different groups of BALB/c mice, once. Additionally, the mice in the control group were given only deionized water. After 14 days of post-IP6-CS-MNPs administration, in a similar way to the untreated mice, the treated mice showed no sign of mortality and abnormalities. However, the serum urea level of mice receiving 2000 mg/kg b.w of IP6-CS-MNPs was significantly higher than the control group (p &lt, 0.05). The mice that received 1000 mg/kg IP6-CS-MNPs showed a significantly higher level of serum alkaline phosphatase (ALP) compared to the control group. However, there were no significant histopathological changes seen in the liver and kidneys of treated mice compared to the untreated group.

Melastoma malabathricum is a well-known herb in Malaysia where it being used in various ways for treatment of different diseases and ailments including skin problems. The study aims to investigate acute and subacute dermal toxicity of ethanolic extract of M. malabathricum leaves following to a single or repeated doses exposure. A total of 30 female Sprague-Dawley rats were grouped into 5 groups (n = 6 per group) for both acute and subacute toxicity study. The duration for each study was determined at 14 days for acute toxicity and 28 days for subacute toxicity. The rats were topically applied with the plant extract at three different doses; 2.5%, 5.0% and 10.0% on the shaved area of dorsal skin. For acute toxicity study, rats in all three groups received single application of the extract on the first day of the experimental period, while rats in subacute toxicity study were topically applied with the extract once daily for 28 days. Throughout the respective 14-day and 28-day study periods, all rats were monitored for any changes in their physical appearance and behavioural patterns that might develop due to toxic effects of the plant. There were no mortality or abnormal physical appearance, and physiological and behavioural changes observed in all rats in both studies. Body weights, kidney and liver weights, and both haematology and serum biochemistry results showed no significant (p > 0.05) differences between all groups in both studies. All of the findings were supported by normal macroscopic and microscopic architectures of liver, kidneys and skin of all rats applied topically with the extract. This study suggests that topical application of M. malabathricum leaf ethanolic extract at 2.5%, 5% and 10% does not induce acute and subacute adverse effects on the skin or systemic toxic reactions in rats.

Despite the ubiquity of microplastics (MPs) in aquatic environments and their proven ability to carry a wide variety of chemicals, very little is known about the impacts of virgin or contaminant-loaded MPs on organisms. The primary aim of this study was to investigate the impacts of virgin or phenanthrene (Phe)-loaded low-density polyethylene (LDPE) fragments on a suite of biomarker responses in juvenile African catfish (Clarias gariepinus). Virgin LDPE (50 or 500µg/L) were preloaded with one of two nominal Phe concentrations (10 or 100µg/L) and were exposed to the fish for 96h. Our findings showed one or both Phe treatments significantly increased the degree of tissue change (DTC) in the liver while decreased the transcription levels of forkhead box L2 (foxl2) and tryptophan hydroxylase2 (tph2) in the brain of C. gariepinus. Exposure to either levels of virgin MPs increased the DTC in the liver and plasma albumin: globulin ratio while decreased the transcription levels of tph2. Moreover, MPs modulated (interacted with) the impact of Phe on the DTC in the gill, plasma concentrations of cholesterol, high-density lipoprotein (HDL), total protein (TP), albumin, and globulin, and the transcription levels of fushi tarazu-factor 1 (ftz-f1), gonadotropin-releasing hormone (GnRH), 11 β-hydroxysteroid dehydrogenase type 2 (11β-hsd2), and liver glycogen stores. Results of this study highlight the ability of virgin LDPE fragments to cause toxicity and to modulate the adverse impacts of Phe in C. gariepinus. Due to the wide distribution of MPs and other classes of contaminants in aquatic environments, further studies are urgently needed to elucidate the toxicity of virgin or contaminant-loaded MPs on organisms.

Lawsonia inermis is one of the most significant plants used in traditional medicine. However, many details of the dermal toxicity of L. inermis remain unknown. The objective of this study is to evaluate the in vivo acute and sub-acute dermal toxicity of ethanolic extract of L. inermis leaves. In acute experiment, a total of 20 rats were divided into four groups of five rats. A total of 30 rats were divided into five groups of six rats for the sub-acute experiment. The extract at a single dose of 2000 and 5000 mg/kg of body weight did not produce treatment-related signs of toxicity or mortality in all rats tested during the 14-day observation period. However, in a repeated dose 28-day study, the application of 500, 1000 and 2000 mg/kg of body weight/day of leaves extract revealed no significant change (p > 0.05) in bodyweight, haematological and biochemical parameters compared with the control group. Similarly, gross pathology and histopathology examinations of liver, kidneys, and skin did not reveal any morphological alteration. Overall, the results show that the close application of L. inermis leaves extract did not have any critically dangerous impact on rats. Subsequently, the concentrate may be employed for pharmaceutical plans.

The gram-negative bacterium Pasteurella multocida serotype B: 2 is an important veterinary and opportunistic human pathogen that causes deplorable welfare problems and colossal economic production losses in bovine and other animal species throughout the world. Pasteurella multocida type B: 2 and its LPS are associated with Haemorrhagic Septicaemia (HS) in mice as animal model. Sixty healthy Balb c mice were placed in twelve plastic cages each one containing five mice. The mice were divided into three major groups (A, B and C). Group A is the control group (n = 10) and these were inoculated with 0.4 mL of PBS pH 7.4 orally. The treatment groups (B; n = 25 and C; n = 25) were inoculated with P. multocida type B: 2 and lipopolysaccharide respectively. The mice in group B and C were further divided into five subgroups. The subgroups were designated based on the graded doses as B101, B103, B105, B107 and B109 for Pasteurella multocida and C101, C103, C105, C107 and C109 for LPS respectively. The mice were observed for 120 h post-inoculation. The concentration of IL-1I² was significantly higher (p

Colorectal cancer (CRC) is the third most common malignancy in males and the second most common cancer worldwide. Chronic colonic inflammation is a known risk factor for CRC. Cocoa contains many polyphenolic compounds that have beneficial effects in humans. The objective of this study is to explore the antioxidant properties of cocoa in the mouse model of azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis-associated cancer, focusing on the activation of Nrf2 signaling. Mice were treated with AOM/DSS and randomized to receive either a control diet or a 5 and 10% cocoa diet during the study period. On day 62 of the experiment, the entire colon was processed for biochemical and histopathological examination and further evaluations. Increased levels of malondialdehyde (MDA) were observed in AOM/DSS-induced mice; however, subsequent administration of cocoa decreased the MDA. Enzymatic and nonenzymatic antioxidants, such as superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase, were decreased in the AOM/DSS mice. Cocoa treatment increases the activities/levels of enzymatic and nonenzymatic antioxidants. Inflammatory mediators, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, were elevated during AOM/DSS-induction, and treatment with 5 and 10% cocoa effectively decreases the expression of iNOS and COX-2. The NF-E2-related factor 2 and its downstream targets, such as NQO1 and UDP-GT, were increased by cocoa treatment. The results of our study suggest that cocoa may merit further clinical investigation as a chemopreventive agent that helps prevent CAC.

Background: Brucellosis in male goats is characterized by arthritis, orchitis and epididymitis, which may induce infertility. Nevertheless, these lesions were categorized as chronic while acute lesions had not been described. This study investigates the histopathological and immuno histochemistry reactions in organs of bucks acutely infected by Brucella melitensis. Results: Only testis and prepuce of acutely infected bucks showed significantly severe histological lesions. Other internal organs had mild to moderate lesions. However, positive immunohistochemistry stainings were observed in organs except the bulbourethral gland. There was a significant positive correlation between the distribution of B. melitensis and IHC intensity but no significant correlation between the IHC intensity and histopathology lesions. Conclusion: The results indicate that acute brucellosis did not lead to clinical presentation, although B. melitensis was well distributed in various organs of bucks.

Boid inclusion body disease (BIBD) is a viral disease of boid snakes believed to be caused by reptarenavirus belonging to the family Arenaviridae. Unlike most mammalian arenaviruses, the reservoir host for reptarenavirus is still unknown. In this study, the pathological responses were evaluated in a mouse model for a period of 28 days. Blood and tissue samples (lung, liver, spleen, heart, kidney and brain) were collected for evaluation of hematology, biochemistry, histopathology and oxidative enzyme levels at six time points (1, 3, 7, 14, 21 and 28 days), after viral infection (2.0 × 106 pfu/mL) in the infected and normal saline in the control groups. An initial increase (p < 0.05) in white blood cell (WBC), neutrophil and lymphocyte counts were observed in the infected group at day 3 post infection, and a decline (p < 0.05) on day 7 and 4 post infection. Significant (p < 0.05) increases in alanine transaminase (ALT), aspartate transaminase (AST), creatinine, total protein and globulin levels were also observed in the infected group. An increased (p < 0.05) level of hydrogen peroxide, total antioxidant capacity (TAC), superoxide dismutase (SOD) activity and catalase activity (CAT) were frequently observed on different days in the infected group. The MDA activity was increased (p < 0.05) in the infected group on day 7 and 14. Histopathological changes observed in the liver, kidney, spleen, brain and lungs were mainly associated with degeneration, necrosis and infiltration of lymphocytes. Viral counts were low on days 7 and 14 but surged in both the liver and spleen on day 21 and 28. This study has shown that reptarenavirus replicates in mammalian host and induces oxidative stress. Furthermore, the resultant hematobiochemical and histopathological changes observed in infected mice were similar to what has been reported in mammarenavirus infections. This suggests that rodents may serve as potential reservoir hosts for reptarenavirus.

A comparative study was conducted to determine the nutrient composition of different feed formulations and its effect on nutritional related blood profile in breeder goats. Eighteen female adult goats, Capra aegagrus hircus (Boer cross) were selected and allocated into three different groups (n = 6) and assigned according to different feed formulations. Blood sampling was conducted before and after fourth weeks of feeding trial and analysed for selected parameters. Proximate analysis of diets was done and revealed comparable value of crude protein and crude fat. Glucose, total protein and calcium level in serum showed significant difference between groups (P-value 0.05). Fatty acid analysis of feed and serum showed polyunsaturated fatty acid was higher than satureated fatty acid. Overall, diet 3 shows the highest value of all blood parameters analysed in this study. Hence, it is very important to provide animal with nutrient according to production stages as sufficent protein and energy will improve the animal performance in terms breeding efficacy and quality of animal products. This study indicates that different feed formulation affects certain nutritional related blood profile in goat.

Balanced nutrition is essential for health and performance of livestock. It is best when feed is formulated according to production stages of goat. However, as the goat grows, it may undergo transition of different feed formulation to meet its daily requirement. The transition may affect the performance of animals as well as it is a stressful condition. In this study, body weight gain and feed intake were used as indicators on performance, whereas hematological stress leucograms were used as indicator of stress parameter during implementation of different feed formulation among does. Eighteen adult does with similar body condition score and body weight approximately 20 kg were selected and divided into three groups equally (n = 6) and fed with different feed formulations, which were formulated using local feed source in the farm for a period of four weeks. Body weight was measured before implementation, on week 2 and week 4, blood sampling was conducted before implementation and during week 4 for comparison and feed intake was measured every day. All data were statistically analyzed using SPSS. Does fed with Diet 3 showed highest body weight gain and feed intake, followed by Diet 2 and Diet 1. There was significant difference (P 0.05) as compared to normal value. Neutrophils:lymphocytes ratio for all groups were within normal range and no significant difference (P>0.05). The implementation of feed formulation according to production stages is recommended as it increases performance and prevents from nutritional stress.

Boid inclusion body disease (BIBD) is a viral disease of boids caused by reptarenavirus. In this study, tissue from naturally infected boid snakes were homogenized and propagated in African Monkey kidney (Vero) and rat embryonic fibroblast (REF) cells. Virus replication was determined by the presence of cytopathic effect, while viral morphology was observed using transmission electron microscopy. Viral RNA was amplified using RT-PCR with primers specific for the L-segment of reptarenavirus; similarly, quantification of viral replication was done using qPCR at 24–144 h postinfection. Viral cytopathology was characterized by cell rounding and detachment in both Vero and REF cells. The viral morphology showed round-to-pleomorphic particles ranging from 105 to 150 nm which had sand-like granules. Sanger sequencing identified four closely associated reptarenavirus species from 15 (37.5 %) of the total samples tested, and these were named as follows: reptarenavirus UPM-MY 01, 02, 03, and 04. These isolates were phylogenetically closely related to the University Helsinki virus (UHV), Boa Arenavirus NL (ROUTV; BAV), and unidentified reptarenavirus L20 (URAV-L20). Comparison of deduced amino acid sequences further confirmed identities to L-protein of UHV, L-polymerase of BAV and RNA-dependent RNA polymerase of URAV-L20. Viral replication in Vero cells increased steadily from 24 to 72 h and peaked at 144 h. This is the first study in South East Asia to isolate and characterize reptarenavirus in boid snakes with BIBD.

The quality characteristics, bioactive phytochemicals, volatile compounds, and antioxidant capacities of virgin avocado oil extracted using a couple of green methods, namely, subcritical CO₂ extraction (SCO₂) and ultrasound‐assisted aqueous extraction (UAAE), are compared with the oil extracted using the conventional solvent extraction. Results indicate the quality properties of avocado oil are unaffected by extraction methods. The total phenolic content of avocado oil is in the range of 111.27–130.17 mg GAE/100 g and the major phytosterol is β‐sitosterol (1.91–2.47 g kg⁻¹). Avocado oil extracted using SCO₂ exhibits two to four times greater levels of α‐ and γ‐tocopherols than solvent extraction and UAAE. The volatile components associated with nutty and grassy flavors are only detected in avocado oil extracted under low‐temperature extraction conditions such as SCO₂ and UAAE. Based on the antioxidant capacity tests, avocado oil obtained by SCO₂ exhibits the strongest antioxidant capacity compared with solvent extraction and UAAE. Practical Applications: Virgin avocado oil extracted using subcritical CO₂ method exhibits superior levels of tocopherols content and antioxidant capacity than ultrasound‐assisted aqueous and conventional solvent methods. Subcritical CO₂ is a promising oil extraction technique to obtain solvent‐free avocado oil with potential uses in functional foods and health care products. Virgin avocado oil is extracted using subcritical CO₂ and ultrasound‐assisted aqueous techniques. The properties of the oil are compared with conventional solvent‐extracted oil. The percentage of antioxidant activity decreases in the order of SCO₂‐extracted oil > solvent‐extracted oil > UAAE‐extracted oil.