Identification of sheep eimerian parasites, Eimeria crandallis and Eimeria faurei, employing microscopic and molecular tools.

Coccidiosis, caused by apicomplexan Eimeria species, is a protozoan disease that affects various species of wild and domestic animals. However, data available on Eimeria diversity in ruminants in Saudi Arabia is meagre. Therefore, this study was designed to investigate some eimerian parasites infecting sheep (Sawakni and Harrie breeds) using microscopy and molecular methods for the first time in Saudi Arabia. Twenty-four fecal samples were collected from sheep farms. Based on the floatation technique, eimerian oocysts were observed in 8 of the 24 (33.33%) fecal samples. The coccidian-positive samples were subjected to fecal culture in a shallow layer of 2.5% potassium dichromate (K2 Cr2 O7 ). Detected eimerian oocysts were described micromorphometrically as the basis for traditional oocyst identification. Morphologically, the sporulated oocysts were similar to those of sheep eimerian parasies; Eimeria faurei and Eimeria crandallis. PCR products from the two eimerian species detected from Sawakni and Harrie breeds were sequenced and were found to be distinct from each other with mutations at five positions. One of them clustered with E. crandallis with 99.8%-100% identity with sequences available in GenBank. E. crandallis was obtained from two Sawakni sheep and two Harrie sheep. The other sequences grouped with E. faurei with 99.8% identity with the only sequences available in GenBank. E. crandallis was detected from both Sawakni and Harrie breeds whereas E. faurei was detected only from Sawakni sheep. The findings of this study have implications for the importance of morphometric identification with advanced molecular tools to confirm the identities of sheep Eimeria species and to address the taxonomic study of this eimeriid parasite at the species level.

Identification of Eimeria acervulina (Apicomplexa: Eimeriidae) infecting the broiler chicken Gallus gallus domesticus through morphology and molecular analyses.

Coccidiosis is an intestinal protozoan disease that affects the poultry industry worldwide. The severity of this disease varies depending on the identity of the infectious agents. Therefore, this study was carried out to identify the Eimeria species that affect broiler chickens, Gallus gallus domesticus, through morphological and molecular phylogenetic analyses. Twenty-five faecal samples were collected from the broiler chickens in a commercial poultry farm in Riyadh (Saudi Arabia). Using the floatation technique, faeces were examined microscopically for the Eimeria occurrence. Identification of Eimeria species was performed based on morphological criteria and molecular tools (DNA amplification for the partial small subunit ribosomal RNA (18S rRNA), internal transcribed spacer (ITS)-1, and mitochondrial cytochrome c oxidase I (COI) genes. In this study, 32% (8 out of 25) of collected samples were found to be positive for coccidiosis. After sporulation in potassium dichromate (K2Cr2O7), the sporulated oocysts were observed as ovoid and measured 18.37-23.19 µm (19.87) long and 15.07-18.67 µm (16.46) wide, with the anterior location of a polar granule and absence of micropyle. These Eimeria oocysts were assumed to size and shape characteristics of Eimeria acervulina. Molecular analysis was conducted on the sequences of the polymerase chain reaction products from the three genes studied (18S rRNA, ITS-1, and COI). At the three genes, results showed that the resultant sequences clustered with E. acervulina from different regions confirming morphological description. This study highlighted the importance of molecular techniques to detect avian Eimeria species more than the traditional morphology-based tool to optimise the appropriate anticoccidial strategies for long-term control in the studied area.

Molecular characterization of ticks and tick-borne piroplasms from cattle and camel in Hofuf, eastern Saudi Arabia.

The aims of the present study were to characterize ticks infesting the dromedary camel and cattle in Hofuf, Eastern Saudi Arabia and to determine the piroplasms that they may harbor. DNA was extracted from ticks, collected from camels and cattle, using commercial kits and subjected to polymerase chain reaction using specific primers for the amplification of ticks and piroplasms DNA. The cytochrome oxidase subunit I mitochondrial gene (COI) was used for characterization of ticks whereas partial 18S rRNA gene (18S rRNA) was used for piroplasms characterization. Ticks were genetically identified as Hyalomma dromedarii and Hyalomma anatolicum. Both cattle and camel in Hofuf, were found to be infested with both species. Both ticks identified as H. dromedarii and H. anatolicum from camels and cows showed 100% identity to COI sequences from the same species available in GenBank. Only Theileria annulata DNA was amplified from both H. anatolicum and H. dromedarii infesting cattle. None of the ticks collected from camels revealed DNA of piroplasms. T. annulata DNA was reported for the first time from Hofuf and the role of both H. anatolicum and H. dromedarii as potential vectors for this parasite in cattle in Saudi Arabia has been documented for the first time.

Morphological and molecular characterization of Aspiculuris tetraptera (nematoda: Heteroxynematidae) from Mus musculus (rodentia: Muridae) in Saudi Arabia.

Aspiculuris tetraptera a pinworm of mice, is an important parasite in institutions with mice colonies for both research and teaching purposes. Infection with this parasite has impact on biomedical research. This is likely due to the availability of the parasite's eggs in the environment, therefore can easily be transmitted and infection is generally asymptomatic. No information regarding the prevalence, morphology or phylogeny is available on A. tetraptera from Saudi Arabia. A group of 50 laboratory mice were investigated for the presence of A. tetraptera. Worms were described morphologically and molecular characterization was attempted using 18S rRNA and Cytochrome Oxidase Subunit I genes. The prevalence of A. tetraptera infestation in the laboratory mice examined was found to be 46%. Morphological description indicated that the worms belong to A. tetraptera and this was confirmed by molecular characterization. Both regions studied have shown that the worm under investigation grouped with A. tetraptera. 18S rDNA sequences obtained in the present study showed high identity with sequences from A. tetraptera while Cytochrome c Oxidase subunit I gene (COI) sequences showed intraspecific variation resulted into two haplotypes from the isolates in the present study. A. tetraptera was recorded for the first time from Saudi Arabia. Molecular characterization has shown, based on the COI sequences, that the Saudi isolates of A. tetraptera are distinct.

Delayed Hemolytic Transfusion Reaction in a Patient with Sickle Cell Disease: Case Report.

BACKGROUND: Blood transfusion is a key treatment of sickle cell disease (SCD) complications. Delayed hemolytic transfusion reaction (DHTR) is a delayed reaction, that occurs days to weeks following a transfusion, characterized by mild anemia and/or hyperbilirubinemia and is one of the serious complications of blood transfusion. The symptoms of DHTR resemble those of vaso-occlusive crisis secondary to SCD leading to difficulty or delaying in diagnosis of DHTR. DHTR may lead to multiple organ failure and death. CASE REPORT: A 31-year-old female patient with a known case of SCD presented to our ER in King Fahad hospital Hofuf in the Kingdom of Saudi Arabia, with a history of generalized body ache, exertional dyspnoea, headache and easy fatigability for a few days on a background history of episodic hospital admissions for SCD, but she was admitted 3 times over the previous 6 months and received 6 units of packed red blood cells (PRBCs). The last blood transfusion was 18 days earlier. She was sick and her Hb level was 4.5 g/dL with positive Coombs test and positive alloantibodies, diagnosed as DHTR. We treated her with prednisolone tablets 1 mg/kg daily, intravenous immunoglobulins, 0.4 gm/kg daily for 5 days, and rituximab 500 mg IV every week for 4 weeks. Her Hb level raised up to 8.2 g/dL and she was discharged in good condition. CONCLUSION: Identifying risk factors for DHTR by history and presentation is urgently needed in order to risk stratify the transfusion regimen. It is important to avoid additional transfusions in these patients if possible because these may exacerbate the hemolysis and worsen the degree of anemia.

Genotyping of Clostridium perfringens Isolates from Domestic Livestock in Saudi Arabia.

The present study was undertaken to confirm the genetic identity of Clostridium perfringens isolates from domestic livestock in Saudi Arabia and to characterize the genes encoding to alpha, beta, epsilon, and iota (α-, ß-, ε-, and ι-) toxins. C. perfringens were confirmed in 104 out of 136 isolates on multiplex polymerase chain reaction using specific primers amplifying genes related to toxins produced by C. perfringens. Genes encoding α-toxins were detected in 104 samples. Of the isolates, 80.8% were diagnosed as type A, 15.4% as type D, 2.9% as type C, and 0.96% as type B. None of the isolates has genes encoding iota (ι-) toxin. All isolates investigated yielded enterotoxin (cpe) products and none yielded ß2 (cpb2-toxin) or NetB products. PLC gene sequences encoding α-toxin showed >96.7% similarity. Isolates which had α-toxins as well as enterotoxin (cpe) are regarded as type F. Phylogenetic analysis using maximum likelihood analysis yielded two clades, and the majority of the isolates were in one group while only two isolates clustered on the second clade. Within the Kingdom of Saudi Arabia strains, 54 variable positions and 23 polymorphic amino acids were noticed. Isolates with ε- and ß-toxins were variable and were found to be close to those published for C. perfringens. ETX gene sequences encoding ε-toxins were found to be related to CPE sequences.

The Effect of Serum Ferritin Level on Gonadal, Prolactin, Thyroid Hormones, and Thyroid Stimulating Hormone in Adult Males with Sickle Cell Anemia.

BACKGROUND: Sickle cell anaemia (SCA) is an inherited hemoglobinopathy resulting in sickling of erythrocytes that cause micro-vascular obstruction leading to acute complications and chronic organ damage. Adults with SCA have endocrine complications and metabolic alterations. The aim of this study was to assess the association between gonadal and thyroid hormones with iron indices and to explore the potential association between serum ferritin levels and sex hormones in adult males with sickle cell disease in New Prince Saud Bin Jalawy Hospital (NPSBJH) in Hofuf city (Eastern region of Saudi Arabia) where there are many patients with SCA. METHODS: A cross-sectional analytical study was carried out in the Haematology Clinic at NPSBJH in 2018. A total of eighty (80) male patients with sickle cell anaemia were included in this study and were divided into two groups according to serum ferritin level. Group I (G-I): Included 40 male patients with high serum ferritin level and group II (G-II): included 40 male patients with normal serum ferritin level. RESULTS: There was a significant difference in height/cm between GI and Gil, P value= 0.006. Serum ferritin was significantly higher in GI (P value= 0.000), and serum TIBC was significantly higher in G-II. (P value= 0.022). Testosterone level was significantly higher in G-II (P value= 0.018). Luteinizing hormone (LH) was significantly higher in group I (P-value 0.019). There was a significant relation between serum ferritin level in G-I and the following: serum iron, TIBC, serum testosterone, LH, prolactin, free T3 and free T4. CONCLUSION: Adult males with SCD with high serum ferritin level were shorter than adult males with SCD who had normal serum ferritin level and had a significant lower level of serum testosterone and significant high level of LH and this was most likely due to endocrine dysfunction secondary to high ferritin level and iron overload.

Seroprevalence of Toxoplasma gondii in household and stray cats of Riyadh, Saudi Arabia.

This study was undertaken to determine the seroprevalence of Toxoplasma gondii in cats in the area of Riyadh, Saudi Arabia. We examined 200 serum samples collected from stray and household cats for T. gondii antibodies by ELISA. The overall seroprevalence was 26%. Seroprevalence was significantly higher (p < 0.05) in stray cats (39%) compared with household cats (13%). The prevalence in male and female cats was 31.4% and 20.4%, respectively. The seroprevalence increased with age and was higher in cats over 6 years of age (43%) as opposed to cats less than 4 years old (33%). Seropositivity varied according to the breed. The highest was recorded among cats of American breed (38.5%), followed by Persian (27%), Himalayan (21%), Bengali (11.5%), and Siamese (2%). Antibodies were not reported from the Turkish breed. Overall seroprevalence among cats did not vary significantly with season or with the localities within the Riyadh municipality. We also examined 100 faecal samples from stray and household cats by flotation technique, which revealed an overall prevalence of 12% of T. gondii oocycts.

The Beneficial Effect of Cape Gooseberry Juice on Carbon Tetrachloride- Induced Neuronal Damage.

OBJECTIVE: Cape gooseberry (Physalis peruviana L.) belongs to the Solanaceae family. Physalis has many medicinal properties however, the beneficial effect of physalis in protecting against neurotoxins has not yet been evaluated. This experimental study investigated the protective effect of physalis juice against the oxidative damage induced by carbon tetrachloride (CCl4) in the rat brain. METHODS: The degrees of protection by physalis in brain tissues were evaluated by determining the brain levels of lipid peroxidation, nitric oxide, glutathione content and antioxidant enzyme activities (superoxide dismutase, catalase, glutathione-S-transferase, glutathione peroxidase and glutathione reductase), after CCl4) induction in the presence or absence of physalis. Adult male albino Wistar rats were divided into 4 groups, Group I served as the control group, Group II was intraperitoneally treated with 2 ml CCl4)/kg bwt for 12 weeks, Group III was supplemented with physalis juice via the drinking water for 12 weeks, Group IV was supplemented with physalis juice and was intraperitoneally injected weekly with CCl4). RESULTS: Treatment with CCl4) was significantly associated with a disturbance in the oxidative status in the brain tissues; this was marked by a significant (p<0.05) elevation in the lipid peroxidation and nitric oxide levels with a concomitant reduction in glutathione content compared to the control, along with a remarkable reduction in antioxidant enzymes. The administration of physalis along with CCl4) juice significantly (p<0.05) alleviated the changes in enzymatic antioxidant activity when compared to the CCl4) treated group. Furthermore, physalis juice supplemention inhibited apoptosis, as indicated by the increase of Bcl-2 immunoreactivity in brain tissue. CONCLUSION: Our results suggest that physalis juice could be effective in preventing neurotoxicity and the neuroprotective effect of physalis might be mediated via antioxidant and anti-apoptosis activities.

Olive oil oleuropein has anti-breast cancer properties with higher efficiency on ER-negative cells.

Breast cancer constitutes a major health problem for women worldwide. However, its incidence varies between populations and geographical locations. These variations could be diet-related, since there are several carcinogenic compounds in the modern diet, while natural products contain various anti-cancer elements. Several lines of evidence indicate that, in addition to their clear preventive effect, these compounds could also be used as therapeutic agents. In the present report we have shown that oleuropein, a pharmacologically safe natural product of olive leaf, has potent anti-breast cancer properties. Indeed, oleuropein exhibits specific cytotoxicity against breast cancer cells, with higher effect on the basal-like MDA-MB-231 cells than on the luminal MCF-7 cells. This effect is mediated through the induction of apoptosis via the mitochondrial pathway. Moreover, oleuropein inhibits cell proliferation by delaying the cell cycle at S phase and up-regulated the cyclin-dependent inhibitor p21. Furthermore, oleuropein inhibited the anti-apoptosis and pro-proliferation protein NF-κB and its main oncogenic target cyclin D1. This inhibition could explain the great effect of oleuropein on cell proliferation and cell death of breast cancer cells. Therefore, oleuropein warrants further investigations to prove its utility in preventing/treating breast cancer, especially the less-responsive basal-like type.

Oleuropein induces anti-metastatic effects in breast cancer.

Breast cancer causes death due to distant metastases in which tumor cells produce matrix metalloproteinase (MMP) enzymes which facilitate invasion. Oleuropein, the main olive oil polyphenol, has anti-proliferative effects. This study aimed to investigate the effect of oleuropein on the metastatic and anti-metastatic gene expression in the MDA human breast cancer cell line. We evaluated the MMPs and TIMPs gene expression by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) in treated and untreated cells. This study demonstrated that OL may induce anti-metastatic effects on human breast cancer cells. We found that TIMP1,-3, and -4 were over-expressed after all periods of incubation in treated cancer cells compared to untreated cells, while MMP2 and MMP9 genes were down-regulated, at least initially. Treatment of breast cancer cells with oleuropein could help in prevention of cancer metastasis by increasing the TIMPs and suppressing the MMPs gene expressions.

Arginine 16 Glycine Polymorphism in ß2-Adrenergic Receptor Gene is Associated with Obesity, Hyperlipidemia, Hyperleptinemia, and Insulin Resistance in Saudis.

Background. Several studies have shown an association between codon 16 polymorphism of the ß2AR gene and obesity. Methods. We studied the association between Arg16Gly polymorphism and obesity and its influence on anthropometric parameters, lipids, insulin resistance and leptin in Saudi individuals. The study group included 329 individuals (males: 109 and females: 220). Metabolic parameters, including glucose, lipids, insulin, and leptin were analyzed and anthropometric parameters including waist and hip circumference, waist/hip (W/H) ratio, and body mass index (BMI) were measured and HOMA-IR was calculated. Genotyping was conducted by DNA sequencing of 353 bp fragments, carrying the Arg16Gly polymorphic site. Results and Conclusion. Overweight and obese subjects had a significantly higher frequency of Gly16 (0.375 and 0.38, resp.) compared with normal-weight subjects (0.200). In addition, subjects carrying Gly16 allele regardless of their BMI had greater waist and hip circumference, W/H ratio, plasma lipids, leptin, glucose level, and insulin resistance as judged from the HOMA-IR, compared to those with the wild-type allele. The findings of this study show a significant association between the Arg16Gly polymorphism in ß2AR gene and the development of insulin resistance, overweight, and obesity in Saudi populations with an influence on the levels of lipid and leptin.

Bisphenol A induces hepatotoxicity through oxidative stress in rat model.

Reactive oxygen species (ROS) are cytotoxic agents that lead to significant oxidative damage. Bisphenol A (BPA) is a contaminant with increasing exposure to it and exerts both toxic and estrogenic effects on mammalian cells. Due to limited information concerning the effect of BPA on liver, this study investigates whether BPA causes hepatotoxicity by induction of oxidative stress in liver. Rats were divided into five groups: The first four groups, BPA (0.1, 1, 10, 50 mg/kg/day) were administrated orally to rats for four weeks. The fifth group was taken water with vehicle. The final body weights in the 0.1 mg group showed a significant decrease compared to control group. Significant decreased levels of reduced glutathione, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, glutathione reductase and catalase activity were found in the 50 mg BPA group compared to control groups. High dose of BPA (50 mg/kg) significantly increased the biochemical levels of ALT, ALP and total bilirubin. BPA effect on the activity of antioxidant genes was confirmed by real time PCR in which the expression levels of these genes in liver tissue were significantly decrease compared to control. Data from this study demonstrate that BPA generate ROS and reduce the antioxidant gene expression that causes hepatotoxicity.

Redescription of Eimeria dorcadis Mantovani, 1966 (Apicomplexa: Eimeriidae) from the dorcas gazelle (Gazella dorcas) in Saudi Arabia.

Eimeria dorcadis Mantovani, 1966 is redescribed from dorcas gazelle (Gazella dorcas (L.)) from Saudi Arabia. Oocysts were detected in 7 out of 22 faecal samples (32%) using floatation method. The sporulated oocysts are cylindrical, slightly flattened at the micropylar pole, measure in average 32 x 19 microm (27-36 x 16-24 microm), length/width ratio being 1.7 (1.5-2.1). Oocyst wall is 1.2 microm thick, smooth, double-layered; outer layer is slightly thicker, light blue in colour; inner layer brownish, with micropyle in the inner layer and apparently continual outer one, measures 2.2 microm, but lacks a micropylar cap. The sporocyst elongate-ellipsoidal, measures 14 x 8 microm (12-17 x 6-9 microm), length/width ratio being 1.8, with sporocyst residuum as circular compact, coarse, refractile granules. Stieda body is present, while substieda body is absent. Sporozoites banana-shaped, measure 11 x 2.5 microm, each with a large spheroidal refractile body at the wider pole. Sporulation time is 2-3 days at 25 +/- 2 degrees C.

Gastrointestinal parasites and their prevalence in the Arabian red fox (Vulpes vulpes arabica) from the Kingdom of Saudi Arabia.

The gastrointestinal parasites and prevalence of infestation in the Arabian red fox Vulpes vulpes arabica Thomas, were investigated at the King Khalid Wildlife Research Centre (KKWRC) in Thumamah, Riyadh Province, Saudi Arabia. Faecal samples were collected from 58 wild caught foxes while under anaesthesia and examined for gastrointestinal parasites stages. Male and female foxes were infected with three major groups of parasites; cestodes, nematodes, protozoa as well as an acanthocephalan. Faecal analyses revealed that 22 foxes (37.9%) were infected with two different Isospora spp. and three (5.2%) with an undescribed Eimeria sp., 12 (20.7%). Nine individuals (15.5%) harboured hookworms, (Trichosomoides sp.), two (3.5%) were infected with Trichuris sp. (probably Trichuris vulpes) and one individual (1.7%) with Taenia sp. (probably Taenia hydatigena). Carcasses of five male and three female foxes were necropsied. Four of the necropsied carcasses yielded Ancylostoma caninum, two each harboured Pterygodermatitis affinis, T. vulpes and Macracanthorhynchus catalinus, in six foxes Joyeuxiella echinorynchoides was found. Five and four foxes were infected with T. hydatigena and Diplopylidium nölleri, respectively. The possible role of the Arabian red fox as an intercalary host essential for the life cycle of Trichosomoides sp., common to the Libyan jird, Meriones libycus, in particular and the importance of this species as a vector for zoonotic infections and in the spread of other parasites to wild and domestic animals in general is discussed.

Coagulation profile and platelet parameters of the Arabian sand gazelle (Gazella subgutturosa marica): comparison with humans and camels.

During March 2009, we evaluated the hemostatic profile and platelet indices of 18 Arabian sand gazelles (Gazella subgutturosa marica) and compared the results with those from humans and camels (Camelus dromedarius). Gazelles and camels had shorter activated partial thromboplastin times, lower proconvertin and higher antihemophilic factor coagulation activity, and plasma fibrinogen levels than humans. Prothrombin time was longer in sand gazelles and shorter in camels than it was in humans. Plasma thromboplastin component, Stuart factor, and plasma thromboplastin antecedent were similar in gazelles, humans, and camels, whereas the platelet count of the sand gazelle was significantly higher than it was for camels and humans.

The efficacy of ivermectin and levamisole against natural Nematodirus spathiger infection in the Arabian sand gazelle (Gazella subgutturosa marica) and the Arabian mountain gazelle (Gazella gazella) in Saudi Arabia.

The efficacy of two broad spectrum anthelmintics (Ivermectin and Levamisole) against the nematode Nematodirus spathiger in the reem (Arabian sand gazelle) and idmi (Arabian mountain gazelle) at King Khalid Wildlife Research Centre (KKWRC) was determined. Gazelles of both species naturally infected with N. spathiger were each divided into 3 groups. One group was treated with Ivermectin, the other group with Levamisole and the last group was left as untreated control. Faecal egg counts were employed to determine the efficacy of the two drugs against N. spathiger infection in both gazelle species. Reduction in arithmetic mean faecal egg counts achieved by Ivermectin and Levamisole in the reem gazelles was 94% and 89.3%, respectively. The reduction in the idmi gazelles was 97.2% and 96.4%, respectively. Hence, both anthelmintics appeared to be more effective in the idmi gazelles than in the reem gazelles, however, there was no significant difference in the faecal egg reduction tests in both animal species.