The High Potency of Green Synthesized Copper Nanoparticles to Prevent the Toxoplasma gondii Infection in Mice.

PURPOSE: Nowadays, due to the lack of an effective vaccine to prevent the toxoplasmosis, chemotherapy with the combination of pyrimethamine and sulfadiazine is considered as the "gold standard" treatment for toxoplasmosis. Recent reports have exhibited that these synthesized chemical drugs are associated with some serious side effects. The present study aims to evaluate the prophylactic effects of copper nanoparticles (CuNPs) green synthesized by Capparis spinosa fruit methanolic extract alone and combined with atovaquone against chronic toxoplasmosis induced by the Tehran strain of Toxoplasma gondii in mice METHODS: Mice were then orally administrated with CuNPs at the doses of 2 and 4 mg/kg/day and in combined with atovaquone 50 mg/kg for 14 days. Male BALB/c mice were divided into two seven groups include C1 (non-treated non-infected); C2 (treated with normal saline); C3 (Infected mice treated with atovaquone 100 mg/kg/day); Ex1 (treated with CuNPs 2 mg/kg/day); Ex2 (treated with CuNPs 4 mg/kg/day); Ex3 (treated with CuNPs 2 mg/kg/day + atovaquone 50 mg/kg/day); Ex3 (treated with CuNPs 4 mg/kg/day + atovaquone 50 mg/kg/day). On the 15th day, the mice were infected with the intraperitoneal inoculation of 20-25 tissue cysts from the Tehran strain of T. gondii. The mean numbers of brain tissue cysts and the mRNA levels of IL-12, IFN-γ, and inducible nitric oxide synthase (iNOS) in mice of each tested group were measured. RESULTS: CuNPs were green synthesized by C. spinosa methanolic extract. Scanning electron microscopy showed that the particle size of CuNPs was 17 and 41 nm with maximum peak at the wavelength of 414 nm. The mean number of T. gondii tissue cysts in mice of tested groups of Ex1, Ex2, Ex3, and Ex4, significantly decreased as a dose-dependent response compared with control group. Moreover, in similar to the control group C3, no T. gondii tissue cysts was observed in mice of experimental group Ex3 and Ex4. The mRNA levels of IFN-γ, IL-12, and iNO was measured in mice of all tested groups. The mRNA levels of IFN-γ, IL-12, and iNO was increased in all mice of experimental groups in comparison with the control group C2; however, a significant enhancement was detected in mRNA level of IFN-γ, IL-12, and iNO in the tested groups of Ex3 and Ex4 when compared with control group C3. CONCLUSION: The obtained results revealed the high potency of CuNPs alone and combined with atovaquone to prevent toxoplasmosis in mice. Although, the prophylactic effects of CuNPs and other properties, such as improved cellular immunity and low toxicity, are positive topics; however, more studies are required to approve these findings especially in clinical settings.

Molecular characterization of Leishmania species from stray dogs and human patients in Saudi Arabia.

Leishmania major and Leishmania tropica cause cutaneous leishmaniasis in humans and dogs in several parts of the world, with a large number of cases recorded in the Middle East. However, when they occur in sympatry, the role of each species of Leishmania in the epidemiology of cutaneous leishmaniasis (CL) is not clear. To assess the frequency and to identify the species of Leishmania that infect humans and stray dogs in Riyadh and Al-Qaseem (Saudi Arabia), 311 stray dogs and 27 human patients who were suspected for Leishmania infection were examined for CL by a nested polymerase chain reaction (nPCR). Seven (25.9%) out of 27 human patients scored positive for Leishmania spp. (i.e., L. major in five patients from Riyadh and L. tropica in two patients from Al-Qaseem). Out of 311 dogs, five (1.6%) were infected by L. tropica. Data herein presented demonstrate the occurrence of L. tropica in dogs and humans in Saudi Arabia, as well as the occurrence of L. major in humans.

Fe3O4@piroctone olamine magnetic nanoparticles: Synthesize and therapeutic potential in cutaneous leishmaniasis.

BACKGROUND: In recent years, magnetic nanoparticles (NMP) as novel materials have been widely used for biomedical, diagnostic and therapeutic purposes like microbial infection therapy. The purpose of this study is to synthesize PO coated iron oxide magnetic nanoparticles (Fe3O4@PO NPs) and their anti-leishmanial effects in vitro and in vivo against cutaneous leishmaniasis. METHODS: Fe3O4 magnetic nanoparticles were synthesized by the coprecipitation of Fe2 + and Fe3 + ions and used as a nanocarrier for the production of Fe3O4@PO NPs. The in vitro antileishmanial effects of PO-coated Fe3O4 NPs and Fe3O4 NPs (10-200 µg/mL) was determined against the intracellular amastigotes of Leishmania major (MRHO/IR/75/ER) and, then, examined on cutaneous leishmaniasis induced in male BALB/c mice by L. major. The rate of infectivity, production of nitric oxide (NO), and cytotoxic activates of Fe3O4 NPs and Fe3O4@PO NPs on J774-A1 macrophage cells were determined. RESULTS: The size scattering of the Fe3O4 NPs and Fe3O4@PO NPs were in the range among 1-40 and 5-55 nm, respectively. The obtained IC50 values were 62.3 ± 2.15 µg/mL, 31.3 ± 2.26 µg/mL, and 52.6 ± 2.15 µg/mL for the Fe3O4 NPs and Fe3O4@PO NPs, and MA, respectively. The results revealed that the mean number of parasites and the mean diameter of the lesions was considerably (p < 0.05) decreased in the infected mice treated with Fe3O4 NPs and Fe3O4@PO NPs. The Fe3O4 NPs and Fe3O4@PO NPs significantly (p < 0.05) prompted the production of NO as a dose-dependent manner. The promastigotes pre-incubated in Fe3O4 NPs and Fe3O4@PO NPs at the concentration of 5 µg/mL had the ability to infect only 41.7% and 28.3% of the macrophages cells. The selectivity index of greater than 10 for Fe3O4 NPs and Fe3O4@PO NPs showed its safety to the J774-A1 macrophage cells and specificity to the parasite. CONCLUSION: The results of this survey indicated the high potency of Fe3O4@PO NPs to inhibit the growth of amastigote forms of L. major as well as recovery and improvement CL induced by L. major in BALB/c mice without significant cytotoxicity. The results also indicated that, although the possible anti-leishmanial mechanisms of Fe3O4@PO NPs have not been clearly understood, however, the triggering of NO may be considered as one of the possible anti-leishmanial mechanisms of these nanoparticles. However, additional studies, in particular in clinical contexts, are mandatory.

Chemical Composition, Apoptotic Activity, and Antiparasitic Effects of Ferula macrecolea Essential Oil against Echinococcus granulosus Protoscoleces.

BACKGROUND: Today, the present protoscolicidals used to minimize the serious risks during hydatid cyst surgery are not completely safe and have various adverse side effects. The present study aimed to evaluate the chemical composition and apoptotic activity of Ferula macrecolea essential oil (FMEO) as well as its in vitro and ex vivo protoscolicidal effects against hydatid cyst protoscoleces. METHODS: Gas chromatography/mass spectrometry (GC/MS) analysis was performed to determine the chemical composition of FMEO. Protoscoleces of hydatid cysts were collected from liver fertile hydatid cysts of infected sheep and were then treated with various concentrations of the essential oil (75, 150, and 300 µL/mL) for 5-60 min in vitro and ex vivo. Then, by using the eosin exclusion test, the viability of the protoscoleces was studied. The caspase-3-like activity of the FMEO-treated protoscoleces was also evaluated through the colorimetric protease assay Sigma Kit based on the manufacturer's instructions. RESULTS: According to GC/MS, the main constituents of the essential oil were terpinolene (77.72%), n-nonanal (4.47%), and linalool (4.35%), respectively. In vitro, the maximum protoscolicidal activity of FMEO was observed at the concentrations of 150 and 300 µL/mL, such that 100% of the protoscoleces were killed after 30 and 20 min of exposure, respectively. Based on the obtained findings, the results demonstrate that FMEO required a longer time to kill protoscoleces ex vivo; after 12 min of exposure to FMEO, only 13.4% of the protoscoleces remained alive. After 48 h of the treatment of protoscoleces, FMEO, in a dose-dependent manner and at doses of 75, 150, and 300 µL/mL, induced the activation of the caspase enzyme by 24.3, 35.3, and 48.3%, respectively. CONCLUSIONS: Our findings demonstrate the potent protoscolicidal effects of FMEO in vitro and ex vivo; however, further studies are required to assess the safety and the efficiency of FMEO as a promising scolicidal agent in a preclinical model and clinical setting.

A report on tick burden and molecular detection of tick-borne pathogens in cattle blood samples collected from four regions in Saudi Arabia.

Babesiosis, theileriosis and anaplasmosis are among the most commonly reported tick-borne diseases in cattle and are associated with significant economic losses. Through the present study the researchers aimed to report the presence of various pathogens that cause babesiosis, theileriosis and anaplasmosis in cattle collected from different provinces in Saudi Arabia and to report their phylogenetic relationship. A total of 362 blood samples of cattle along with ticks that were present on the cattle were collected from four regions (Riyadh, Al-Kharj, Al-Hasa and Al-Qassim) of Saudi Arabia. Blood samples were screened by polymerase chain reaction (PCR) for the presence of various Babesia, Theileria and Anaplasma species by amplification of their 18S rRNA and/or 23S rRNA genes. A total of 541 ticks were collected and identified from the cattle. These included Hyalomma anatolicum, Hyalomma dromedarii, Hyalomma impeltatum, Hyalomma excavatum, Rhipicephalus annulatus and Rhipicephalus turanicus. Regarding tick-borne pathogens, the overall prevalence was 1.9 % (7/362) for Theileria annulata, (2/362) 0.6 % for Theileria and (21/362) 5.8 % for Anaplasma ovis. Four of the cattle were found to be co-infected with more than one pathogen (1.1 %). We did not detect any Babesia species in the blood of the studied cattle. Prevalence of the Theileria and Anaplasma species was highest in cattle that resided in Riyadh, followed by cattle from Al-Hasa and Al-Qassim. Representative amplified partial-gene sequences of T. annulata (GenBank accession numbers MK826137-39) and A. ovis (GenBank acc. no. MK 880224) were submitted to GenBank. The presence of ticks on cattle was found to be associated with a high prevalence of Theileria spp. (P = 0.02) and Anaplasma ovis (P < 0.001). We report novel genotypes of T. annulata and A. ovis from cattle in Saudi Arabia and we recommend that molecular surveys are undertaken throughout the country to address the prevalence and geographical distribution of tick-borne infections for their effective diagnosis and treatment.

Seroprevalence, Risk Factors and Molecular Identification of Bovine Leukemia Virus in Egyptian Cattle.

Bovine leukemia virus (BLV) is distributed worldwide and affects dairy cattle causing severe economic losses. The BLV has been serologically reported in Egypt, but few studies have evaluated its associated risk factors and genetic classification. Therefore, this study assessed risk factors associated with BLV infection and identified the genetic diversity of the Egyptian strain. The study was conducted on 500 dairy cattle distributed in four Governorates located in Northern Egypt. Overall, the seroprevalence of BLV infection among Egyptian dairy cattle was 18.2%. The grazing cattle in the losing house system had higher odds for BLV seropositivity, and bad practice such as the use of a single needle or one plastic glove for more than one animal was considered a significant risk factor for BLV infection. Besides, the sequencing and phylogenetic analysis for one Egyptian BLV strain was performed, and the obtained results confirmed the clustering of Egyptian BLV strain into genotype-1. The assessment of associated risk factors for BLV infection and determination of its genetic classification are essential to implement an effective control program.

Protective effects of thymoquinone against acrylamide-induced liver, kidney and brain oxidative damage in rats.

Acrylamide (AA), an industrial monomer, may cause multi-organ toxicity through induction of oxidative stress and inflammation. The antioxidant properties of thymoquinone (TQ), an active constituent of Nigella sativa, have been established before. The aim of the current study was to assess the protective effects of TQ against AA-induced toxicity in rats. Forty-eight male Wistar rats were divided into six groups each of eight rats. The first group acted as a negative control and received normal saline. Groups II and III were administered TQ orally at doses of 10 and 20 mg/kg b.wt., respectively, for 21 days. The four group received AA (20 mg/kg b.wt.) for 14 days. The five and six groups were given TQ at either dose for 21 days, starting seven days before AA supplementation (for 14 days). Acrylamide intoxication was associated with significant (p < 0.05) increases in serum levels of liver injury biomarkers (alanine transferase, aspartate transferase, and alkaline phosphatase), renal function products (urea, creatinine), DNA oxidative damage biomarker (8-oxo-2'-deoxyguanosine), and pro-inflammatory biomarkers (interleukin-1ß, interleukin-6, and tumor necrosis factor-α). Moreover, AA intoxication was associated with increased lipid peroxidation and nitric oxide levels, while reduced glutathione concentration and activities of glutathione peroxidase, superoxide dismutase, and catalase in the liver, kidney, and brain. TQ administration normalized AA-induced changes in most serum parameters and enhanced the antioxidant capacity in the liver, kidney, and brain tissues in a dose-dependent manner. In conclusion, the current experiment showed that TQ exerted protective and antioxidant activities against AA-induced toxicity in mice.

Ticks and associated pathogens in camels (Camelus dromedarius) from Riyadh Province, Saudi Arabia.

BACKGROUND: Camel production in Saudi Arabia is severely affected by various diseases and by inadequate veterinary services. Ticks and tick-borne pathogens (TBPs) affect the health and wellbeing of camels consequently diminishing their productivity and performances. In addition, camels may act as hosts for TBPs (e.g. Anaplasma phagocytophilum) causing diseases in humans. The current study aimed to determine the prevalence of ixodid ticks and molecularly investigate the associated pathogens in camels from Saudi Arabia. METHODS: Blood and tick samples were collected from camels (n = 170) in Riyad Province of Saudi Arabia. Ticks were morphologically identified, and blood of camels were molecularly screened for apicomplexan (i.e. Babesia spp., Theileria spp., Hepatozoon spp.) and rickettsial parasites (i.e. Ehrlichia spp. and Anaplasma spp.). RESULTS: Of the 170 camels examined, 116 (68.2%; 95% CI: 60.9-75.1%) were infested by ticks with a mean intensity of 2.53 (95% CI: 2.4-2.6). In total of 296 ticks collected, Hyalomma dromedarii was the most prevalent (76.4%), followed by Hyalomma impeltatum (23.3%) and Hyalomma excavatum (0.3%). Of the tested animals, 13 (7.6%; 95% CI: 4.3-12.8%) scored positive to at least one TBP, with Anaplasma platys (5.3%; 95% CI: 2.7-9.9%) being the most prevalent species, followed by Anaplasma phagocytophilum, Anaplasma sp., Ehrlichia canis and Hepatozoon canis (0.6% each; 95% CI: 0.04-3.4%). None of the camels were found to be co-infected with more than one pathogen. All samples tested negative for Babesia spp. and Theileria spp. CONCLUSIONS: The present study reveals the occurrence of different tick species and TBPs in camels from Saudi Arabia. Importantly, these camels may carry A. phagocytophilum and A. platys, representing a potential risk to humans.

Fucoidan Ameliorates Oxidative Stress, Inflammation, DNA Damage, and Hepatorenal Injuries in Diabetic Rats Intoxicated with Aflatoxin B1.

The current study was carried out to evaluate the ameliorative effect of fucoidan against aflatoxicosis-induced hepatorenal toxicity in streptozotocin-induced diabetic rats. Sixty-four Wister albino male rats were randomly assigned into eight groups (8 rats each) that received normal saline, fucoidan (FUC) at 100 mg/kg/day orally for 4 weeks, streptozotocin (STZ) at 50 mg/kg/i.p. single dose, STZ plus FUC, aflatoxin B1 (AFB1) at 50 µg/kg/i.p. after one month of the beginning of the experiment for 2 weeks, AFB1 plus FUC, STZ plus AFB1, or STZ plus AFB1 and FUC. Injection of rats with STZ induced hyperglycemia. Rats with STZ-induced diabetes, with or without AFB1 intoxication, had significantly elevated activities of serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase, and levels of serum urea, creatinine, cholesterol, 8-oxo-2'-deoxyguanosine, interleukin-1ß, interleukin-6, and tumor necrosis factor-α. In addition, these rats exhibited increased lipid peroxidation and reduced glutathione concentration and activities of superoxide dismutase, catalase, and glutathione peroxidase enzymes in the hepatic and renal tissues. In contrast, administration of FUC to diabetic rats, with or without AFB1 intoxication, ameliorated the altered serum parameters, reduced oxidative stress, DNA damage, and inflammatory biomarkers, and enhanced the antioxidant defense system in the hepatic and renal tissues. These results indicated that FUC ameliorated diabetes and AFB1-induced hepatorenal injuries through alleviating oxidative stress, DNA damage, and inflammation.

Fucoidan protects against subacute diazinon-induced oxidative damage in cardiac, hepatic, and renal tissues.

Fucoidans (FUC) are organic sulfated polysaccharides from natural seaweeds with multiple biological actions. The current study was performed to assess the chemoprotective, antioxidant, and anti-inflammatory effects of FUC from Laminaria japonicum against diazinon (DZN)-induced injuries to rat cardiac, hepatic, and renal tissues. Forty male Wistar rats were assigned into five groups, receiving saline, oral FUC 200 mg/kg/day, subcutaneous DZN 20 mg/kg/day, DZN plus FUC 100 mg/kg/day, or DZN plus FUC 200 mg/kg/day (each treatment was given daily for 4 weeks). Data analysis showed that DZN-intoxicated rats exhibited significantly higher (p < 0.05) serum levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, urea, creatine, creatine kinase, creatine kinase-MB, lactate dehydrogenase, cholesterol, interleukin-6, and tumor necrosis factor-α, as well as lower levels of acetylcholinesterase, compared to control rats. In addition, DZN intoxication was associated with significantly higher (p < 0.05) cardiac, hepatic, and renal tissue concentrations of malondialdehyde and nitric oxide, as well as lower glutathione concentrations, and activities of glutathione peroxidase, superoxide dismutase, and catalase enzymes in comparison to control rats. Treatment with FUC (at 100 or 200 mg/kg/day) ameliorated all the aforementioned alterations in a dose-dependent manner. In conclusion, FUC from Laminaria japonicum ameliorated DZN-induced oxidative stress, pro-inflammatory effects, and injuries to the cardiac, hepatic, and renal tissues. These effects may be related to the antioxidant and anti-inflammatory effects of FUC.

Molecular Survey of Vector-Borne Pathogens of Dogs and Cats in Two Regions of Saudi Arabia.

Dogs and cats play an important role as reservoirs of vector-borne pathogens, yet reports of canine and feline vector-borne diseases in Saudi Arabia are scarce. Blood samples were collected from 188 free-roaming dogs and cats in Asir (70 dogs and 44 cats) and Riyadh (74 dogs), Saudi Arabia. The presence of Anaplasma spp., Bartonella spp., hemotropic Mycoplasma spp., Babesia spp., and Hepatozoon spp. was detected using a multiplex tandem real-time PCR. PCR-positive samples were further examined with specific conventional and real-time PCR followed by sequencing. Dogs from Riyadh tested negative for all pathogens, while 46 out of 70 dogs (65.7%) and 17 out of 44 cats (38.6%) from Asir were positive for at least one pathogen. Positive dogs were infected with Anaplasma platys (57.1%), Babesia vogeli (30%), Mycoplasma haemocanis (15.7%), and Bartonella henselae (1.4%), and cats were infected with Mycoplasma haemofelis (13.6%), Candidatus Mycoplasma haemominutum (13.6%), B. henselae (9.2%), and A. platys (2.27%), all of which are reported for the first time in Saudi Arabia. Co-infection with A. platys and B. vogeli was detected in 17 dogs (24.28%), while coinfections were not detected in cats. These results suggest that effective control and public awareness strategies for minimizing infection in animals are necessary.

Molecular Detection of Leishmania spp. in Skin and Blood of Stray Dogs from Endemic Areas of Cutaneous Leishmaniasis in Saudi Arabia.

BACKGROUND: Dogs can act as reservoirs of canine leishmaniasis, caused by Leishmania species. The aims of this study were to determine the prevalence of canine leishmaniasis using a PCR technique among stray dogs living in three provinces of Saudi Arabia, Riyadh, Al-Ahsa Oasis and Al-Qaseem, where the disease is endemic; and to identify and document different Leishmania to species levels. METHODS: This cross-sectional investigation was conducted, from Mar 2016 to Apr 2018, in three parts of Saudi Arabia: Central province (Riyadh), Eastern province (Al-Ahsa Oasis) and Al-Qaseem province. Blood samples were collected from 526 dogs; 40 presented cutaneous nodules so were suspected clinically of cutaneous leishmaniasis. Biopsy tissue collections and parasite cultures were performed. A generic kDNA was performed using different primers for Leishmania differentiation. RESULTS: All blood samples were negative for Leishmania infantum infection by molecular analysis, though forty dogs had thick cutaneous lesions in different parts of their body. Four dogs' skin lesions were associated with dermatitis, splenomegaly and lymphadenomegaly. Parasite culture was used to diagnose cutaneous leishmaniasis, identifying 31/40 (77.5%) positive samples. Overall, of 526 samples, the prevalence of L. major and L. tropica was found to be 4% and 1.9%, respectively. Gender and age had a significant effect on Leishmania prevalence: (P=0.0212 and 0.0357), respectively. CONCLUSION: This was the first molecular study of dog leishmaniasis from Saudi Arabia of dogs confirmed to have cutaneous leishmaniasis. Further epidemiological and molecular investigations of domestic and wild canine infections with L. major, L. tropica and L. infantum in endemic and nonendemic areas of Saudi Arabia are required, for leishmaniasis control.

Rhipicephalus sanguineus sensu lato from dogs and dromedary camels in Riyadh, Saudi Arabia: low prevalence of vector-borne pathogens in dogs detected using multiplexed tandem PCR panel.

Despite the global distribution of the brown dog tick, Rhipicephalus sanguineus (Latreille, 1806) sensu lato (s.l.), limited information exists about their identity from the Arabian Peninsula. Ticks from free roaming urban dogs and dromedary camels in Riyadh, Saudi Arabia were morphologically identified, confirmed with scanning electron microscopy and characterised at mitochondrial DNA (cox1, 12S rDNA and 16S rDNA). A total of 186 ticks were collected from 65 free roaming dogs (n = 73) and 84 dromedary camels (n = 113). Morphologically, 5.9% (11/186) were R. sanguineus s.l. and Hyalomma spp. (93.5%, 174/186). From within R. sanguineus s.l., the presence of Rhipicephalus cf. camicasi Morel, Mouchet et Rodhain, 1976 (1 dog, 2 camels) and Rhipicephalus turanicus Pomerantsev, 1936 (1 camel) is reported. The examined R. cf. camicasi form a sister group to R. sanguineus s.l. tropical lineage at all DNA markers. Dogs were parasitised by Hyalomma dromedarii Koch, 1844 (n = 59), Hyalomma impeltatum Schulze et Schlottke, 1930 (n = 1), Hyalomma excavatum Koch, 1844 (n = 2), Hyalomma turanicum Pomerantsev, 1946 (n = 1) and Hyalomma rufipes Koch,1844 (n = 1). DNA from dog blood (n = 53) from Riyadh confirmed a low prevalence of canine vector-borne pathogens that does not exceed 5.7% for Babesia spp., Mycoplasma spp., Anaplasma platys, Hepatozoon canis and Ehrlichia canis using multiplexed tandem PCR (MT-PCR) and diagnostic PCR. Low prevalence of R. sanguineus s.l. on dogs likely contributed to the low level of canine vector-borne pathogens in Saudi Arabia. We demonstrate that dogs in the central Arabian Peninsula are more commonly parasitised by Hyalomma spp. than R. sanguineus s.l.

Species Diversity and Seasonal Distribution of Hard Ticks (Acari: Ixodidae) Infesting Mammalian Hosts in Various Districts of Riyadh Province, Saudi Arabia.

Hard ticks are among the most important blood sucking arthropods that transmit pathogens to humans and animals. This study was designed to determine prevalence, mapping, geographical distribution, and seasonal activity of hard tick species infesting the most common domestic and wild mammals in various districts of Riyadh Province, Saudi Arabia, during the period January to December 2017. In total, 10,832 adult hard ticks were collected from the bodies of 8,435 animals belonging to 18 different mammalian species. The ticks were preserved in 70% alcohol and microscopy was used to identify species. Two genera, Hyalomma and Rhipicephalus, were identified, comprising 10 species of hard ticks, with Hyalomma comprising 68.3% and Rhipicephalus comprising 31.7% of species. The most common species on domestic mammalian hosts was Hyalomma dromedarii (Koch 1844) (39.9%) followed by Rhipicephalus turanicus (Pomerantsev, Matikashvili & Lotosky 1936) (34.9%), whereas on wild mammalian hosts Rhipicephalus sanguineus (Latreille 1806) was by far the most prevalent species (83.0%). However, ticks were most abundant during May through July (36.0%) in the studied areas, and tick intensity and abundance differed among seasons. Our results provide information for human and animal health service managers, as well as governmental authorities, to gain a better understanding of hard ticks infesting mammalian hosts in Riyadh Province, Saudi Arabia, which can help improve prevention and control of tick-borne diseases, especially during outbreaks.

Potential role of dogs as sentinels and reservoirs for piroplasms infecting equine and cattle in Riyadh City, Saudi Arabia.

Canine tick-borne diseases have been considered emerging and re-emerging threats, given their increasing global prevalence. In this molecular survey, we aimed to detect and identify common tick-borne pathogens in dogs from Riyadh city in Saudi Arabia. Initially, the study included 36 dogs visiting private veterinary clinics. PCRs targeting the 18S ribosomal RNA gene (rDNA) of haemoparasites (Babesia, Theileria and Hepatozoon) and the 16S rDNA of Anaplasmataceae were performed. The results showed that 26 (72.2%) dogs were infected by some of the haemoparasites under investigation. The sequencing analysis of the amplicons confirmed the infections due to two parasite species Theileria equi and Theileria velifera. Further examination of guard dogs kept in the horse stables of the Riyadh Municipality revealed that the majority of the tested dogs (65.2%: 30 out of 46) were infected with either of the parasites. In addition, the genotypes of all the parasites in these dogs were identical to those of the parasites in the dogs from the veterinary clinics. Thus, it can be concluded that dogs are infected with these haemoparasites and serve as a reservoir for both T. equi and T. velifera in the study area; however, the clinical implication of this finding is to be studied.

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.

Molecular detection of equine trypanosomiasis in the Riyadh Province of Saudi Arabia.

We conducted a cross-sectional study to detect trypanosome infections of horses and donkeys in the Riyadh Province of Saudi Arabia. DNA was extracted from blood samples collected from 368 horses and 142 donkeys, and subjected to universal first ribosomal internal transcribed spacer region (ITS1)-PCR followed by Trypanosoma evansi species-specific RoTat1.2-PCR. The universal ITS1-PCR revealed T. evansi infection in horses ( n = 12; 3.3%) and donkeys ( n = 4; 2.8%). There was no significant effect of sex or age on the prevalence of trypanosomiasis in horses or donkeys. Application of the RoTat1.2-PCR revealed that the RoTat1.2 VSG gene was absent from the positive ITS1-PCR samples of 3 horses and 1 donkey. This discrepancy could be explained by the circulation of T. evansi type B in Saudi Arabia; however, this suspicion requires confirmation.

Detecting early signs of heat and drought stress in Phoenix dactylifera (date palm).

Plants adapt to the environment by either long-term genome evolution or by acclimatization processes where the cellular processes and metabolism of the plant are adjusted within the existing potential in the genome. Here we studied the adaptation strategies in date palm, Phoenix dactylifera, under mild heat, drought and combined heat and drought by transcriptomic and metabolomic profiling. In transcriptomics data, combined heat and drought resembled heat response, whereas in metabolomics data it was more similar to drought. In both conditions, soluble carbohydrates, such as fucose, and glucose derivatives, were increased, suggesting a switch to carbohydrate metabolism and cell wall biogenesis. This result is consistent with the evidence from transcriptomics and cis-motif analysis. In addition, transcriptomics data showed transcriptional activation of genes related to reactive oxygen species in all three conditions (drought, heat, and combined heat and drought), suggesting increased activity of enzymatic antioxidant systems in cytosol, chloroplast and peroxisome. Finally, the genes that were differentially expressed in heat and combined heat and drought stresses were significantly enriched for circadian and diurnal rhythm motifs, suggesting new stress avoidance strategies.

A field study on the anthelmintic resistance of Parascaris spp. in Arab foals in the Riyadh region, Saudi Arabia.

BACKGROUND: In the last decade, Parascaris spp. resistance to anthelmintics has been recorded in many countries. In Saudi Arabia, there are limited data available on Parascaris spp. resistance to anthelmintics. OBJECTIVE: To determine the current status of ivermectin, abamectin and praziquantel combined, and fenbendazole resistance to Parascaris spp. in horses in Saudi Arabia. METHODS: Three hundred and forty-one foals from eleven different farms were examined by faecal egg count (FEC). The foals were all Arab horses aged 17.2 ± 4.5 (SD) months. Ivermectin (n = 46 foals), abamectin and praziquantel combined (n = 46), and fenbendazole (n = 46) were administered on day 0 and faeces were collected on day 14. The study comprised 41 untreated foals as controls. Animals that have FEC of ≥100 eggs per gram (EPG) were used to measure anthelmintic efficacy. Parascaris spp. populations were considered susceptible when faecal egg count reduction (FECR) was ≥95% associated with a lower 95% confidence limit (LCL) >90%, suspected resistant when FECR ≤90% or LCL <90% and resistant when FECR <90% and LCL <90%. RESULTS: Prevalence of Parascaris spp. infection was 53% (179/341 horses). Anthelmintic resistance to Parascaris spp. were highest following fenbendazole (55% of farms and 65% of foals) and to a lower extent following ivermectin or the combination of abamectin and praziquantel which comprised 27% of farms (and 46% of foals) and 18% of farms (and 10% of foals), respectively. CONCLUSION: These data indicate that anthelmintics-resistant Parascaris spp. populations are present on horse farms in Saudi Arabia.