Evaluation of oxidative stress and antioxidant status, serum trace mineral levels and cholinesterases activity in cattle infected with Anaplasma marginale.

This study was undertaken to assess the influence of an Anaplasma marginale infection on oxidative stress and antioxidant status, trace elements and cholinesterase as markers of the inflammatory process and biomarkers of oxidative imbalance. An infected group comprised of 35 crossbred Holstein cattle, about 2-3 years old, naturally infected with Anaplasma marginale, were divided into 4 subgroups according to their parasitemia rates (<1%, 1-10%, 10-20%, >20%) and also 10 healthy cattle as control were selected. Blood samples were taken and hematological parameters, activities of antioxidant enzymes including erythrocyte glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD), total antioxidant capacity (TAC), median corpuscularfragility (MCF) as well as acetylcholinesterase (AChE), and serum concentrations of antioxidant trace minerals (copper, iron, zinc, manganese, and selenium) and butyrylcholinesterase (BChE) were determined. In addition, as an index of lipid peroxidation, the level of malondialdehyde (MDA) was measured. The results revealed a significant decrease (P < 0.05) in RBC count, packed cell volume (PCV) and Hb concentration as well as the activities of erythrocyte GSH-Px, SOD, CAT, G6PD, TAC, MCF and AChE and serum concentrations of Cu, Zn, Mn, Se and BchE in the infected cattle. In contrast, significantly increased (P < 0.05) levels of MDA and erythrocyte osmotic fragility as well as serum concentration of iron were recorded in the infected animals. The significant decrease in antioxidant enzyme activities and substantial elevated levels of lipid peroxidation and erythrocyte osmotic fragility associated with the notable increase in parasitemia indicate increased exposure of RBCs to oxidative damage. Furthermore, decrease of cholinesterase in infection by A. marginale can and directly or indirectly lead to increase acetylcholine levels potent anti-inflammatory molecules, thereby inhibiting inflammation.

Molecular verification of transplacental transmission of Theileria lestoquardi in goat.

Ovine and caprine malignant theileriosis (OCMT), a critical condition in small ruminant production, causes lethal infections. In September 2016, a total number of 400 goats of Marghoz breed (the largest population of goat breed in Iran), in northwest of Iran, were examined for approximately 25 pre-partum abortions and 7 goats' mortality for a period of 3 days. A dead goat and her aborted fetus were brought into the Urmia Veterinary Hospital for further diagnostic investigations. The microbiological assessments including direct microscopical examination of the vaginal discharges and placentome with respect to the differential staining (Gram's staining method), conventional pure culturing, fetal abomasal contents, and the liver were negative. Microscopic examinations of blood smears of the goat and the fetus revealed characteristic of Theileria spp. piroplasms and impression smear samples from goat liver and fetal spleen were positive for Theileria Koch blue bodies. DNA analysis was performed using PCR technique and specific primers derived from the nucleotide sequences of 18S rRNA gene of T. lestoquardi; following extraction from blood samples, placentome, goat liver, and spleen of the fetus. The amplified DNA was sequenced afterwards and the corresponding sequence was registered under GenBank accession number MG208059. The sequence alignment showed that the products of PCR had a homology of 99% to known T. lestoquardi sequence registered under accession numbers of KY352037.1, KC778786.1, and JQ917458.1 in the GenBank. To our knowledge, this is a report demonstrating molecular verification of T. lestoquardi transplacental transmission in a neonatal kid of Marghoz breed of goats, its feasible role in induction of perinatal deaths and abortion in goat flocks.

Green synthesis and anthelmintic activity of silver nanoparticles using Morus Alba Fruit extract against different stages of equine strongyles.

The current study was designed to green synthesize silver nanoparticles (GRAgNPs) using Morus alba fruit extract and evaluate their nematicidal effects against strongyle nematodes compared to commercial silver nanoparticles (CAgNPs) in vitro. The nanoparticles were characterized by Ultraviolet-visual absorption spectrography, transmission electron microscopy, and X-ray diffraction. Next, uptake of AgNPs by the first stage larvae (L1), egg hatch inhibition (EHI) and the motility of infectious larvae (L3s), and the ultrastructural analysis of the eggs and worms were conducted. Moreover, some of oxidative/nitrosative stress indicators, including total antioxidant status content (TAC), protein carbonylation (PCO), lipid peroxidation (MDA), and DNA damage were assessed in the homogenized samples of strongyle L3s. We found that the GRAgNPs had spherical shape, 20-30 nm in diameter with rough surface. Following incubation with GRAgNPs at concentrations of 43.40, 21.70 and 10.85 ppm and CAgNPs at concentrations of 43.40 and 21.70, EHI was more than 90%. In addition, concentrations of 43.40 and 21.70 ppm of GRAgNPs led to inhibition of larval motility by more than 90%. The LC50 at 24 h of treatment for GRAgNPs and CAgNPs was determined to be 8.62 and 10.34 ppm, respectively. GRAgNPs and CAgNPs, in a concentration-dependent manner, resulted in the induction of oxidative/nitrosative stress evidenced by decreased TAC levels, and increased levels of MDA and PCO, together with increased DNA damage. The uptake of AgNPs by the L1 larvae revealed that FITC labeled GRAgNPs fluoresced with high intensity largely in the intestinal area. Scanning Electron Microscopy analysis of eggs and larvae revealed that GRAgNPs penetrated the cuticle of larvae, changed the tegmentum, and ultimately killed the worm. In conclusion, GRAgNPs had more robust anthelminthic effects than the standard antiparasitic and CAgNPs. They could be considered as a promising antiparasitic agent.

First report of Hepatozoon felis infection in a domestic cat (Felis catus) in Iran.

A 6-year-old male domestic short-haired cat (Felis catus) was presented with weakness, anorexia, fever, icterus, a painful abdomen, ruffled hair and a tick infestation, and it had no prior surgery. Laboratory analysis revealed left-shifted neutrophils, thrombocytopenia, low albumin content and high serum bilirubin concentration as well as activities of hepatic enzymes including alanine aminotransferase and aspartate aminotransferase. Azotemia and increased serum levels of creatinine and urea were also recorded. In Giemsa-stained blood smear, Hepatozoon gamonts were observed within neutrophils. Species-specific polymerase chain reaction assay was used to amplify an approximately 590 bp fragment of 18S rRNA gene and confirmed Hepatozoon felis infection. The cat was treated with imidocarb dipropionate and doxycycline and recovered completely. Six-month follow-up showed no recurrence. This study reveals the presence of H. felis in Iran and it should be considered in differential diagnosis in febrile and icteric cats. To the authors' knowledge, this is the first description of H. felis infection in a cat in Iran.

Prevalence of intestinal parasites in raw vegetables consumed in Soran city, Kurdistan Region, Iraq.

Intestinal parasitic infections are one of the most important health problems. Although fresh vegetables are considered as a vital element of a healthy diet, they are a potential source of human intestinal parasitic infections during production, collection, transportation, preparation and consumption. The present study was carried out to determine and detecting the parasitological contaminations of fresh vegetables sold at different open-aired markets in Soran city, Iraq. A total of 225 raw vegetable samples were examined by sedimentation technique after washing the samples with normal saline. Of the 225 samples, 109 (48.4%) were microscopically positive for intestinal parasites. Particularly, a high rate of contamination was recorded in cress (Lepidium sativum) with 71.1%. However, the least contaminated sample was lettuce (Lactuca sativa) with 26.6%. The most encountered parasite was Taenia spp. egg (24%), whereas hookworms (4%) were the least detected ones and 34.6% of the samples harbored poly-parasitic contamination. Since, the results emphasize the fresh vegetables are potential sources of transmission for intestinal parasites in central markets of Soran city, strict hygienic measures should always be adopted to avoid parasitic contamination. Untreated sewage and the manure used by greengrocer as fertilizer can be implicated in vegetable contamination.

The Phylogenetic Analysis of Cimex hemipterus (Hemiptera: Cimicidae) Isolated from Different Regions of Iran Using Cytochrome Oxidase Subunit I Gene.

BACKGROUND: Bedbugs are blood feeding ectoparasites of humans and several domesticated animals. There are scarcity of information about the bed bugs population throughout Iran and only very limited and local studies are available. The aim of this study is to assess the phylogenetic relationships and nucleotide diversity using partial sequences of cytochrome oxidase I gene (COI) among the populations of tropical bed bugs inhabiting Iran. METHODS: The bedbugs were collected from cities located in different geographical regions of Iran. After DNA extraction PCR was performed for COI gene using specific primers. Then DNA sequencing was performed on PCR products for the all 15 examined samples. RESULTS: DNA sequencing analysis showed that the all C. hemipterus samples were similar, despite the minor nucleotide variations (within the range of 576 to 697bp) on average between 5 and 10 Single nucleotide polymorphisms (SNPs). Subsequently, the results were compared with the database in gene bank which revealed close similarity and sequence homology with other C. hemipterus from other parts of the world. CONCLUSION: In conclusion, this study has demonstrated the ability of the COI gene to differentiate between the C. hemipterus populations from a few different locations in Iran. The current research is the first report of phylogenetic and genetic species diversity analysis conducted on C. hemipterus in Iran. These results provided basic information for further studies of molecular epidemiology, public health and pest control operators in Iran.

Molecular analysis of pyrethroid resistance in Cimex hemipterus (Hemiptera: Cimicidae) collected from different parts of Iran.

The present study was aimed to assess the bedbugs susceptibility to pyrethroid insecticides using molecular analysis. With the aid of pest control companies, adult bedbugs were collected from various places such as hotels, residential houses, and industrial buildings in seven cities highly crowded with domestic and foreign tourists in Iran from May 2016 to August 2017. Bedbugs were colonized in the laboratory to evaluate their resistance to pyrethroid using insecticide resistance bioassay. Genomic DNA was extracted from susceptible and resistant bedbugs. At first, specie specific primers targeting cytochrome oxidase subunit I (COI) gene was performed to confirm Cimex hemipterus species. Then, kdr-like gene was examined for point mutation using PCR and nucleotide sequencing. Bioassay showed that 11 out of 35 examined bedbugs were resistant to pyrethroids (31.43%; 95.00% confidence interval: 29.48-33.08%). The DNA sequencing showed that all examined bedbugs collected from Tehran province had homozygous V419L kdr-like gene mutations. The level of pyrethroid resistance found in the collected bugs from Tehran province indicated that this phenomenon has already been prevailed in the site and prompts the need to reevaluate the large use of pyrethroids to control the bedbugs.

Hemolytic anemia associated with Trypanosoma theileri in a cow from Kurdistan province, West of Iran.

Various species of Trypanosoma parasites are known to infect several wild and domestic animals worldwide. A 7-year-old Holstein cow from Baneh, Kurdistan province, was examined by a private veterinarian due to anorexia and depression. Physical examination revealed fever, enlarged subscapular lymph node, and pale mucosa. Blood samples were taken for hemato-logical, parasitological, and PCR examination. The large Trypanosoma spp. was microscopically observed in a stained blood smear. Decreased red blood cells (RBCs) count, packed cell volume and hemoglobin concentration were observed through complete blood cell count. Nucleated RBCs were also found in this case. Species-specific PCR assay confirmed T. theileri infection. Treatment was performed subcutaneously with diminazene aceturate. The clinical signs were improved after two days. Two-month follow-up showed no recurrence. In conclusion, T. theileri is characterized by anemia and pyrexia in a cow. To our knowledge, the present case report describes the first molecular evidence of T. theileri in Kurdistan, West of Iran.

Status of oxidative stress, trace elements, sialic acid and cholinesterase activity in cattle naturally infected with Babesia bigemina.

Babesia bigemina infection (also known as Texas fever) is reported as the most prevalent and main causative agent of bovine babesiosis, worldwide. The current study was undertaken to assess indicators of oxidative stress including activities of antioxidant enzymes and total antioxidant capacity (TAC), oxidation status of biomolecules and serum levels of trace elements as well as indicators of inflammation including sialic acid (SA) contents and cholinesterase activity in cattle naturally infected with B. bigemina. An infected group comprised of 20 crossbred Holstein cattle (3-4 years old) were diagnosed to be positive by both microscopy and nested PCR assay. The infected animals were subdivided into two groups according to their parasitemia rates (<20 % and>20 %). Furthermore, 10 healthy cattle were included as the control. The infection caused severe anemia in a parasitemia-burden dependent fashion. The activities of catalase and glucose-6-phosphate dehydrogenase as well as the levels of TAC, zinc, selenium, copper and manganese were significantly decreased as the parasitemia increased, accordingly. However, the activity of superoxide dismutase as well as the levels of malondialdehyde, protein carbonylation, DNA damage and iron, were significantly elevated in a parasitemeia-burden dependent manner. Additionally, glutathione peroxidase activity was significantly elevated with the lower rate of parasitemia, but the higher rate had no significant effect as compared to control. Moreover, total, protein and lipid binding SA contents were significantly increased but the activities of acetylcholinesterase and butyrylcholinesterase were significantly reduced, parasitemia dependently. Conclusively, the infection was remarkably associated with the induction of anemia, oxidative stress and inflammation.

Trypanosoma evansi and "Candidatus Mycoplasma haemolamae" Co-Infection in One-Humped Camel (Camelus dromedarius) from the Northwest of Iran: A Case Report.

A 4-year-old male one-humped camel (Camelus dromedarius) was referred to Veterinary Teaching Hospital of Urmia University, Iran in 2017 with anorexia, weakness, depression and pale mucosa. Decreased red blood cell count, packed cell volume and hemoglobin concentration were detected by complete blood cell count. In Giemsa-stained peripheral blood smears Trypanosoma spp. trypomastigotes scattered between erythrocytic spaces and Mycoplasma-like organisms were observed attached to the surface of erythrocytes. Species-specific PCR assay confirmed T. evansi and Candidatus Mycoplasma haemolamae (CMhl) co-infection. Administration of diminazene aceturate, oxytetracycline 20%, flunixin meglumine and phosphorus-vitamin B12 were not effective in treatment. Hemoplasmosis should be considered as an important differential diagnosis of conditions associated with hemolytic anemia in camel.

A survey of Dictyocaulus arnfieldi (Nematoda) infections in equids in Urmia region, Iran

Dictyocaulus arnfieldi is a lungworm commonly found in equids; however, relatively little is known about it. The aim of the present study was to establish the prevalence of equine lungworms in naturally-infected working and sporting horses, donkeys and mules in rural areas of Urmia, northwest Iran. The fecal samples were collected from 299 working horses, 57 sporting horses, 66 donkeys and 37 mules during the period March 2014 to June 2016. The collected fecal samples were processed within 48 hours following rectal sampling. The larval count/g (LPG) of feces was determined using the Baermann technique. The larvae were identified morphologically, and the arithmetic mean of the LPG was calculated at each sampling point. The overall prevalence of lungworm in all equine species was found to be 19.20%., with specific values of 15.71%, 0%, 31.81% and 24.32% being found in working horses, sporting horses, donkeys and mules, respectively. The working horses, donkeys and mules were also found to be heavily infected with Dictyocaulus arnfieldi and in addition to donkeys and mules, working horses are considered natural hosts for lungworm, and this has effects on both the epidemiology of parasitic infections and performance of the equids.

Assessment of oxidative/nitrosative stress biomarkers and DNA damage in Haemonchus contortus, following exposure to zinc oxide nanoparticles.

Drug resistance in helminth parasites has incurred several difficulties to livestock industry and ranked among the top public health concerns. Therefore, seeking for new agents to control parasites is an urgent strategy. In the recent years, metallic nanoparticles have been considerably evaluated for anthelmintic effects. The current research was conducted to assess possible anthelmintic impacts of zinc oxide nanoparticles (ZnO-NPs) on a prevalent gastrointestinal nematode, H. contortus. Moreover, several biomarkers of oxidative/nitrosative stress and DNA damage were measured. Various concentrations of the nanoparticle were prepared and incubated with the worms for 24 hours. The parasite mobility, mortality rate, antioxidant enzymes activities (SOD, Catalase and GSH-Px), lipid peroxidation, total antioxidant status as well as nitric oxide (NO) contents and DNA damage were determined. ZnO-NPs exerted significant wormicidal effects via induction of oxidative/nitrosative stress and DNA damage. Conclusively, ZnO-NPs can be utilized as a novel and potential agent to control and treatment of helminth parasitic infections.