Evaluation of drug resistance to albendazole and levamisole against lung worms in goat flocks based on fecal larvae count reduction test.

The over-use of anti-parasitic compounds as a method of control has led to insufficient effectiveness and widespread drug resistance worldwide. The aim of this study was to investigate the efficacy of albendazole and levamisole as anti-parasitic agents in a lung worm control program in goat flocks. During 2021 and 2022, a total of 110 goats (age of four months and above) were randomly selected from 11 herds in the north-western region of Iran including Saanen breed (both sexes of the same age). The results indicated that 3.60, 50.80 and 41.90% were respectively infected with Dictyocaulus filaria, Muellerius capillaris and Proto-strongylus rufescens, and generally all the lung parasites in goats of this region were resistant to albendazole and levamisole. Due to clinical importance of D. filaria in goats, the molecular analysis of two samples was also done. Sequencing results showed that the identified parasites were 100% similar to the reference sequences registered in the GenBank®. The results of this research showed low level of these anthelmintics efficacy against Dictyocaulus and Muellerius. Generally, the lung parasites in goats of this region are resistant to albendazole and levamisole. The P. rufescens showed high resistance to these drugs. Totally, it can be concluded that the level of drug resistance varies in different parts of the world; but, the frequencies of drug resistance in different parts of the world are not the same, requiring more studies.

Differential cell analysis in ovine bronchoalveolar lavage fluid: Effect of different sampling methods and seasons.

Background: Bronchoalveolar lavage (BAL) is a method for collecting the cellular and fluid components of the airway surface in the lungs. The assessment of differential cell profiles is potentially valuable in the diagnosis of pulmonary diseases, but there is no information about the normal BAL profiles in the Gezel breed. Aim: This study aimed to characterize the normal cryptologic findings of BAL with Gezel sheep. Methods: Twenty healthy sheep (15 females: 5 males, bodyweight: 55-65 kg) were sedated with xylazine (0.02-0.04 mg/kg IV). Two methods; the transtracheal bronchoalveolar lavage technique and the bronchoscopic bronchoalveolar lavage using a scope were evaluated. Sampling was performed in the summer and winter seasons. Results: Normal value (Mean ± SEM) for total cell, macrophage, lymphocyte, neutrophil, epithelial, and bronchial cells in BAL sampled in summer were (343.75 ± 30.23), ) 24.50 ± 2.62), (2.81 ± 0.51), (1.43 ± 0.88), and (3.12 ± 0.32), respectively. The normal values for the total cell, macrophage, lymphocyte, neutrophil, epithelial, and bronchial cells in BAL sampled in winter were (355.55 ± 37.67), (59.11 ± 4.30), (21.33 ± 3.10), (3.88 ± 1.07), (8.88 ± 3.78), and (6.33 ± 1.44), respectively. Conclusion: No significant change in the percentage of neutrophils was detected between seasons, although the percentages of bronchial and epithelial cells in winter were significantly high (p < 0.05). Except for the mentioned cases, neither the total cell number nor the percentage differential cell populations of BAL changed significantly (p < 0.05) in different sampling methods and seasons. Normal BAL profiles in the Gezel breed were determined and could be used in result interpretations. Also, both sampling methods can be used without significantly affecting the results.

Comparison of the sedative and cardiovascular effects of the combination of acepromazine-clonidine versus acepromazine-xylazine in horses.

The aim of this study was to compare the sedative and cardiovascular effects of the combination of acepromazine-clonidine versus acepromazine-xylazine in horses. Four healthy cross-bred horses were included in the study. They were assigned to two treatments. In treatment I (T1), the animals received xylazine hydrochloride (1.00 mg kg-1) in combination with acepromazine maleate (0.05 mg kg-1) intravenously (IV). In treatment II (T2), the animals received intra-gastric administration of clonidine (0.002 mg kg-1) followed by acepromazine (0.05 mg kg-1; IV) after 60 min. Head height above the ground (HHAG) and echocardiographic indices were evaluated. In T1, recordings were made 5 min before and 5, 15, 30, 60, and 90 min after drug administration. In T2, recordings were made 5 min before clonidine, 55 min after clonidine administration, and then 5, 15, 30, 60, and 90 min after acepromazine injection. Analyses of the data showed there were not significant differences regarding HHAG and echocardiographic indices between two treatments. For sedation of healthy horses, it was concluded that intra-gastric administration of clonidine and IV administration of acepromazine showed similar sedative and cardiovascular effects compared to IV acepromazine-xylazine administration.

Prevalence and Concentration of Mycotoxins in Animal Feed in the Middle East and North Africa (MENA): A Systematic Review and Meta-Analysis.

This study seeks a comprehensive meta-analysis of mycotoxin contaminants in animal feed consumed in the Middle East and North Africa (MENA) region. The obtained articles were reviewed, and 49 articles that investigated the contamination of mycotoxins including aflatoxins (AFs), deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin, fumonisins (FUM), and ochratoxin A (OTA), in feed samples or components of animal feed in the MENA region were selected. The titles of the final articles included in the study were meta-analyzed. Necessary information was extracted and categorized from the articles, and a meta-analysis was performed using Stata software. The highest contamination was in dry bread (80%), and Algeria was the most contaminated country (87% of animal feed), with the most mycotoxins contaminating AFs (47%) and FUM (47%). The highest concentration of mycotoxins in animal feed is related to FUM (1240.01 µg/kg). Climate change, economic situation, agricultural and processing methods, the nature of the animal feed, and improper use of food waste in animal feed are among the most critical factors that are effective in the occurrence of mycotoxin contamination in animal feed in MENA. Control of influential factors in the occurrence of contaminations and rapid screening with accurate identification methods to prevent the occurrence and spread of mycotoxin contamination of animal feed seem important.

Antibacterial interactions of pulegone and 1,8-cineole with monolaurin ornisin against Staphylococcus aureus.

The aim of this study was to investigate the antibacterial interactions of pulegone and 1,8-cineole with monolaurin ornisin against Staphylococcus aureus. The individual and combined antibacterial activities of the compounds were evaluated using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), fractional inhibitory concentration index (FICi), and time-kill methods. Furthermore, the mechanism of the antibacterial action of the compounds was tested by measuring the release of cell constituents. The MIC values of pulegone, 1,8-cineole, nisin, and monolaurin were 5.85 µl/ml, 23.43 µl/ml, 6.25 µg/ml, and 0.031 mg/ml, respectively. A synergistic antibacterial activity (FICi = 0.5) was found between 1,8-cineole and nisin. The time-kill assay showed that the populations of S. aureus exposed to 1,8-cineole, nisin, and their combination were decreased by 5.9, 5.3, and 7.1 log CFU (colony-forming units)/mL, respectively. The combination of 1,8-cineole and nisin also induced the highest release of cell constituents. It was concluded that the combination of 1,8-cineole and nisin could be considered as a novel and promising combination which may reduce the required dose of each antibacterial compound.

Prophylactic effects of Glycyrrhiza glabra root extract on phenylbutazone-induced Equine Glandular Gastric Disease (EGGD).

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs), like phenylbutazone (PBZ), are prescribed to treat pain and inflammation in the equine. However, experimentally their use has been associated with the development of Equine Glandular Gastric Disease (EGGD). To evaluate the prophylactic effects of Glycyrrhiza glabra root extracts (GGRE) on EGGD induction following oral administration of PBZ, 12 donkeys were used. Animals were divided into three equal groups (A, B and C) in a randomized block design. A: placebo, B: PBZ (4.4 mg/kg PO q 12 h) and C: PBZ plus GGRE (17.6 mg/kg PO q 24h). Gastroscopy and blood sampling for CBC, biochemical tests and antioxidant status were performed days 0 and 7. All but 1 animal had no EGGD lesions on day 0, but on day 7, severe EGGD lesions were seen in group B animals (grade 1/4 (× 1), grade 2/4 (× 2), and grade 3/4 (× 1)). Only mild changes were seen in group C animals, the most severe lesions being grade 1/4 lesions in 2 animals. No change was observed in Group A. Hematology and serum antioxidant status did not change significantly in any group. In group B glucose decreased and ALT and ALP increased, while in group C only ALP increased. In conclusion, GGRE reduced the severity of EGGD caused by PBZ. Further, GGRE prevented PBZ induced hypoglycemia and might reduce the possible hepatic injury of PBZ.

Effects of feed restriction and supplementary garlic oil on blood metabolites in ewes.

The effects of garlic oil (GO) on serum fatty acids, lipid profiles and energy-related blood metabolites in feed-restricted fat-tailed ewes were investigated. For this purpose, twenty-eight dry, non-pregnant Makuei ewes (about three years of age; 45.00 ± 2.20 kg of body weight) were assigned randomly to four experimental groups including group 1 receiving basal diet as total mixed ration (TMR) without GO supplementation (control group), group 2 receiving 100% basal diet supplemented with GO (10.00 mg kg-1 BW), group 3 receiving 70.00% basal diet without GO supplementation and group 4 receiving 70.00% basal diet supplemented with GO (10.00 mg kg-1 BW). The main experimental period started eight weeks after performing adaptation and dietary allocations. Feed restriction reduced serum glucose levels along with higher serum levels of non-esterified fatty acids, triacylglycerols, ß-hydroxybutyrate, low-density lipoprotein cholesterol, total cholesterol and very low-density of lipoprotein. Following feed restriction, the serum palmitic and oleic acids concentrations were increased. Garlic oil supplementation had a desirable effect on feed-restricted animals through lowering serum BHB and palmitic and oleic acids concentration and increasing the high-density lipoprotein cholesterol levels in the serum. These findings demonstrated that GO had the potential to reduce body-fat mobilization, thereby lowering the risk factors for disorders associated with negative energy balance in underfed ewes in the periparturient period.

Phylogenetic analysis of the lumpy skin disease viruses in northwest of Iran.

Lumpy skin disease (LSD) is a devastating viral disease of cattle which has recently spread from Africa into the countries of the Middle East. The aim of the present study was to investigate the relationships among lumpy skin disease viruses (LSDV) isolated from different regions of Iran and the origin and spread of these viruses. In this study, a total of 234 blood samples from clinically affected animals from four provinces in the northwest of Iran were screened for LSDV using polymerase chain reaction (PCR). From 80 positive samples for LSDV detected by PCR, the partial P32 gene (759 bp) of 12 isolates were sequenced and phylogenetically analyzed. LSD viruses were grouped in three subclusters with an overall 97.1-100% nucleotide identity. LSDVs isolated from Gilan showed lowest nucleotide identity with the other LSDVs. Four isolates of LSDV including KO-1, EA-1, EA-3, and WA-3 showed 100% similarity with each other and also with the Neethling strain. Phylogenetic analysis indicated that the identified LSDVs were closely related to each other and had high-sequence homology with other LSDV isolates from Africa. It was concluded that LSD outbreak probably occurred in the northwest of Iran by LSDVs entering the country from Iraq and P32 nucleotide sequence information obtained in the present study is a valuable resource in understanding the genetic nature and molecular epidemiology of local LSDV isolates which can be used for future vaccine development based on the circulating strains in the region.

The in vitro effect of Ferula asafoetida and Allium sativum extracts on Strongylus spp.

The high incidence of equine gastrointestinal worms and their increased resistance against anthelmintics has encouraged research into the effectiveness of rational phytotherapy. This study investigates the in vitro anti-parasitic effects of extracts of Ferula asafoetida and Allium sativum, two native plants that are widespread in Iran on Strongylus spp. larvae. Faecal samples were collected from horses, examined by routine parasitology methods and positive samples were used for future examination. After incubation, the third-stage larvae were harvested by the Baermann technique. A hydroalcoholic extract from the plants was used for the antiparasitic study, while tap water was used for controls. Trials for each concentration and control group were performed in three replicates. The results showed that that during the first day of exposure, the hydroalcoholic extract of F. asafoetida at concentration of 10, 50 and 100 mg/ml killed over the 90% of the larvae, and A. sativum extract at concentration of 50 and 100 mg/ml killed over the 95% of larvae (p<0.05). The results obtained from the bioassay showed that two plant extracts have a larvicidal effect on the Strongylus spp. larval stages compared with the control group.

Effect of (R)-(+) Pulegone on Ovarian Tissue; Correlation with Expression of Aromatase Cyp19 and Ovarian Selected Genes in Mice.

OBJECTIVES: Pulegone (PGN) is a monoterpene ketone, whose metabolites exert several cytotoxic effects in various tissues. The present study was conducted in order to evaluate the (R)-(+) PGN-induced alterations in ovarian aromatization, proto-oncogenes and estrogen receptorα ( ERα) and ERß receptors expressions. MATERIALS AND METHODS: In this experimental study, mature albino mice were divided into experimental (received 25 mg/kg, 50 mg/kg and 100 mg/kg PGN, orally for 35 days) and control (received 2% solution of Tween 80 as a PGN solvent, orally) groups. The mRNA levels of Erα, Erß, p53, Bcl-2, and cytochrome p450 (Cyp19) as well as ovarian angiogenesis were analyzed through reverse transcription polymerase chain reaction and immunohistochemical techniques, respectively. Moreover, apoptosis of follicular cells, serum estrogen and progesterone levels and mRNA damage were investigated via using terminal transferase and biotin-16-dUTP staining, electrochemilunescence and fluorescent microscopy methods, respectively. RESULTS: The PGN reduced Erα, Erß and Cyp19 expression at 50 mg/kg and 100 mg/kg doses, while significantly elevating p53 and reducing Bcl-2 expression. Finally, PGN impaired ovarian angiogenesis, increased apoptosis, elevated follicular atresia and reduced serum levels of estrogen and progesterone. CONCLUSIONS: Chronic exposure to PGN (50 mg/kg and 100 mg/kg), severely affects ovarian aromatization, protooncogenes mRNA levels and expression of ERs.

Does Achillea millefolium extracts possess prokinetic effects on the bovine abomasum thourgh M3 muscarinic receptors?

Displacement of the abomasum is a common disease of the gastrointestinal tract in dairy cattle. Abomasal displacement has been associated with abomasal hypomotility. Therefore, it is necessary to identify effective therapeutic agents that stimulate abomasal motility in cattle. Yarrow (Achillea millefolium) is traditionally used as a folk remedy for treatment of human gastrointestinal complaints in the northwest of Iran. This study investigated the effects of A. millefolium extracts on abomasal smooth muscle preparations from healthy cattle. The A. millefolium extracts (3 to3000 mg L-1) contracted the isolated of smooth muscle in a concentration-dependent manner, with an effective threshold concentration of 30 mg L-1 (p < 0.05). The strongest contraction by A. millefolium aqueous extracts in a concentration of 3000 mg L-1 was observed and amounted to 124.90 ± 10.80% of the control treatment. This action was unaffected by pretreatment with hexamethonium and indomethacin, but strongly reduced by verapamil, atropine and 4-DAMP. The inhibiting effect of 4-DAMP and atropine suggesting that the effect of A. millefolium extracts is mediated at least partly by M3 muscarinic acetylcholine receptor. The results suggested that A. millefolium has the potential prokinetic effect that may prevent or alleviate dysfunctions of gastrointestinal motility.

Molecular detection, infection rate and vectors of Theileria lestoquardi in goats from West Azerbaijan province, Iran.

This study was aimed to determine the infection rate and vectors of Theileria lestoquardi in goats from West Azerbaijan province, Iran. A total of 400 blood samples were collected from 40 randomly selected flocks in the study area from June to September, 2014. Out of 400 blood samples examined using microscopic examination, a number of 14 goats (3.50%) were positive for Theileria spp., whereas 25 goats (6.25%) yielded a specific T. lestoquardi SSU-rRNA fragment (235 bp). The prevalence of theileriosis in goats estimated by semi-nested PCR was significantly higher than the prevalence estimated by microscopic examination of the blood smears. The prevalence of Theileria infection in different age and sex groups of goats was not significantly different. The highest and lowest prevalence of Theileria infection was in July (12.00%) and September (2.00%), respectively. A number of 315 adult Ixodid ticks were also collected from naturally infested goats and they were characterized. Out of 315 examined ticks, a number of 37 ticks including Hyalomma marginatum (65.20%), Rhipicephalus turanicus (44.00%), and Dermacentor marginatus (68.70%) were infected by T. lestoquardi. Based on the obtained results, it was concluded that the semi-nested PCR assay based on SSU-rRNA gene is a valuable method for epidemiological investigation of caprine theileriosis. The results showed that H. marginatum, R. turanicus and D. marginatus can be considered as risk factor in the epidemiology of T. lestoquardi.

Planimetric and Biomechanical Study of Local Effect of Pulegone on Full Thickness Wound Healing in Rat.

BACKGROUND: Pulegone as principal component of essential oil, reported to have anti-bacterial, antioxidant and anti-inflammatory properties. The present study was aimed to evaluate wound healing activity of pulegone in a rat model. METHOD: Forty rats were used for excisional and incisional wound healing models. For each model twenty male white Wistar rats were randomly divided into five groups (n = 4) of control (CG), Sham surgery, E1, E2 and E3. Wound size, hydroxyproline content of wound and biomechanical testing were assessed. RESULT: In E2 animals, the wound size was reduced earlier than in E1 and E2 groups (P = 0.035). However, time had significant effect on wound contraction of all wounds. Hydroxyproline contents in the groups CG, sham surgery, E1, E2 and E3 were found to be 51.25 ± 3.40, 58.41 ± 4.62, 68.59 ± 3.53, 86.32 ± 3.18, and 74.26 ± 4.73 mg g-1, respectively. Hydroxyproline contents were increased significantly in E2 compared to E1 and E3 which implied more collagen deposition compared to other experimental groups (P = 0.001). The biomechanical indices, maximum stored energy, stiffness, ultimate strength and yield strength obtained for E2 group were significantly higher than those obtained for E1 and E2 groups (P = 0.002). CONCLUSION: The pulegone showed a reproducible wound healing potential in rats.

Study on the Relationship between Liver Parasitic Infections and Serum Vitamin A and ß-Carotene Status in Cattle.

OBJECTIVE: This study was conducted to evaluate the relationship between serum levels of vitamin A and ß-carotene in the liver of cattle. METHODS: A total number of 150 samples were selected according to the type of parasitic infections of the liver after postmortem examination and confirmation. Parasitic lesions in the liver were subdivided into three major parasites, including Fasciola hepatica, Dicrocoelium dendriticum, and hydatid cysts. Fifty samples were obtained from cattle without any parasitic infection as a control group. Serum levels of vitamin A, ß-carotene concentrations (µg/dL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity were assayed. RESULTS: Naturally infected cattle with D. dendriticum, F. hepatica and hydatid cyst showed lower vitamin A levels. Serum ß-carotene levels were significantly decreased in all groups (p<0.05). ALT and AST activities in animals with parasitic diseases were statistically higher than in control group (p<0.01). A significant negative correlation was determined between the ß-carotene, vitamin A levels and enzyme activities of the liver in all the three types of liver infections (p<0.001). CONCLUSIONS: This study indicated that serum levels of vitamin A and ß-carotene decline was present in cattle with liver parasite infection and vitamin supplements should be supplied.

Antiulcer properties of Glycyrrhiza glabra L. extract on experimental models of gastric ulcer in mice.

Glycyrrhiza glabra L. is used in folk medicine for treatment of stomach disorders including peptic ulcers. The hydroalcoholic extract of Glycyrrhiza glabra L. (HEGG) was evaluated for antiulcerogenic activity and acute toxicity profile in mice. Various doses of HEGG (50-200 mg/kg) were administered orally to animals of different groups. Omeprazole and cimetidine at doses of 30 and 100 mg/kg were used as positive controls, respectively. Stomach was opened along the greater curvature then ulceration index was determined examining the inner lining of stomach. Oral administration of the extract at 1600 mg/kg did not produce toxic symptoms and mortality in mice. 2950 mg/kg was determined as the oral LD50. The HEGG (50-200 mg/kg) showed a significant reduction in ulcer index in HCl/Ethanol-induced ulcer. G. glabra extract (50-150 mg/kg) showed antiulcer activity against indomethacin-induced gastric lesions dose dependently. The extract effectively inhibited formation of gastric lesions induced by ethanol. The extract (200 mg/kg) was more potent than omeprazole (30 mg/kg). HEGG reduced the ulcer index in hypothermic stress induced gastric ulcers in mice and the antiulcer effect was comparable to that of cimetidine. The results indicated that G. glabra hydroalcoholic extract exerted an antiulcergenic effect that could be associated with increase in gastric mucosal defensive factors.

Antidiarrheal activity and acute oral toxicity of Mentha longifolia L. essential oil.

OBJECTIVES: Mentha longifolia L. (Lamiaceae) is an annual herb that is used in the Iranian traditional medicine for treating stomach and intestinal disorders. The purpose of this study was to determine the protective effect of M. longifolia on experimental diarrhea in a rat model. MATERIALS AND METHODS: The antidiarrheal activity of essential oil of M. longifolia (20-80 mg/kg) was investigated against castor oil-induced diarrhea in rats using loperamide as the standard reference drug. In acute toxicity evaluation, rats were orally administrated with single dose of EOML at doses ranging from 10 to 1000 mg/kg. RESULTS: EOML caused a significant (p<0.05) and dose-dependent decrease of gastrointestinal transit, nevertheless, it could not block the inhibitory effect of atropine (0.1 mg/kg). EOML at oral doses of 20 and 80 mg/kg protected the animals against castor oil-induced diarrhea significantly (p<0.05). EOML decreased the intestinal fluid accumulation as indicated by the significantly (p<0.05 to p<0.001) decrease compared to control. The oral LD50 of EOML was found to be 470 mg/kg in rat. CONCLUSION: Since the inhibition of intestinal hyperactivity and hypersecretory are the bases of the treatment of diarrhea, results obtained in the present study suggest that EOML is endowed with antidiarrheal activity. EOML is moderately toxic for oral medication.

Lead and cadmium in raw buffalo, cow and ewe milk from west Azerbaijan, Iran.

In this study, 300 raw buffalo, cow and ewe milk samples from five townships in west Azerbaijan, Iran, were analysed. Lead and cadmium were determined using atomic absorption spectrophotometry. Mean concentration of lead and cadmium in buffalo milk samples was 0.018 ± 0.001 and 0.003 ± 0.001 mg/kg, respectively. Mean concentration of lead and cadmium in cow milk samples was 0.007 ± 0.001 and 0.001 ± 0.001 mg/kg, respectively, and in ewe milk, these mean values were 0.010 ± 0.001 and 0.002 ± 0.001 mg/kg, respectively. Statistical analyses showed that lead and cadmium concentrations in buffalo milk were significantly higher than those in cow and ewe milk. Moreover, the concentration of these heavy metals in ewe milk was significantly higher than that in cow milk. It was also found that concentration of these selected toxic metals in milk increased with increasing age of the animals.

The application of 1,8-cineole, a terpenoid oxide present in medicinal plants, inhibits castor oil-induced diarrhea in rats.

CONTEXT: 1,8-Cineole, a terpene, characterized as a major constituent occurring in the essential oils of several aromatic plants. It is widely used in pharmaceutical industry, as a food additive and for culinary purposes. OBJECTIVE: This study investigates the inhibitory effect of 1,8-cineole on transit time and diarrhea in animal models. MATERIALS AND METHODS: Acute toxicity and lethality of 1-8-cineole was determined by Lork's guidelines. The antidiarrheal effect of 1,8-cineole was investigated by determining the intestinal transit and enterpooling in rats. In all experiments, different doses of 1,8-cineole (20-120 mg/kg), atropine, and loperamide were administered orally. RESULTS: The LD50 of 1,8-cineole for oral administration was estimated to be 1280 mg/kg. 1,8-Cineole (20-120 mg/kg) did not show a significant decrease in small intestine transit (p > 0.05); however, the highest dose displayed a significant decrease in comparison with atropine (p < 0.05). This substance decreased the peristaltic index value to 68 ± 0.36% at a dose of 120 mg/kg compared with the control group (85.22 ± 4.31%) in the castor oil transit test. 1,8-Cineole significantly delayed the onset of diarrhea to -142.33 ± 6.08 min at 120 mg/kg, while the time was 103.66 ± 20.73 min for the control and >240 min for the loperamide. Moreover, 1,8-cineole significantly decreased intestinal fluid accumulation (p < 0.05). CONCLUSIONS: This study demonstrated antispasmodic and antisecretory activities of 1,8-cineole and rationalized the traditional use of the plant containing various levels of this terpene in the treatment of gastrointestinal complains such as diarrhea.

Evaluation of antioxidant status, oxidative stress and serum trace mineral levels associated with Babesia ovis parasitemia in sheep.

Ovine babesiosis is a fatal disease characterized by severe progressive hemolytic anemia. In order to clarify the causal mechanisms implicated in anemia, this study was aimed to assess the antioxidant status and erythrocyte oxidative stress in sheep suffering from ovine babesiosis. Babesia infection was confirmed both with Giemsa's staining blood smears and semi-nested PCR amplified region of 18S rRNA gene. Thirty-eight Iranian sheep, naturally infected with Babesia spp., were considered as the infected group and divided into four subgroups according to parasitemia rates (<1%, 1-2%, 2-3% and >3%), and the same number non-infected animals were selected as the control group. 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 corpuscular fragility (MCF), and serum concentrations of some trace minerals (copper, iron, zinc, manganese, and selenium) were measured. In addition, as an index of lipid peroxidation, the level of malondialdehyde (MDA) was measured. The results revealed a significant decrease (P<0.01) 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 serum concentrations of Cu, Zn, Mn and Se in the infected sheep. In contrast, significantly increased (P<0.01) levels of MDA and erythrocyte osmotic fragility as well as serum concentration of iron were recorded in the infected animals. Overall, the observed remarkable decrease in the antioxidant enzyme activities, median corpuscular fragility and substantial elevated levels of lipid peroxidation associated with the notable increase in parasitemia indicate high exposure of RBCs to oxidative damage in Babesia infected sheep. These results indicate that the disturbed antioxidant defense mechanisms in ovine babesiosis can promote the development of anemia.

PCR-Based Detection of Babesia ovis in Rhipicephalus bursa and Small Ruminants.

This study aimed to assess the prevalence of Babesia ovis infection in adult Rhipicephalus bursa and small ruminants in West Azerbaijan province, Iran. Blood samples were collected from 280 sheep and 122 goats of forty randomly selected flocks. Specific B. ovis fragment was detected in 67 animals (16.7%), of which 52 animals (18.6%) were sheep and 15 animals (12.2%) goats (P < 0.05). Of the 848 R. bursa collected from naturally infested small ruminants and farm dogs, Babesia ovis was detected by PCR in salivary glands of 94 adult ticks. The frequency of B. ovis infection was higher in flocks with tick in comparison with animals without tick (P < 0.05). Positive amplification from blood of ruminants, ticks, oviposition ticks, eggs, and larvae was subjected to restriction digestion with HphI. One RFLP profile was produced. The PCR-RFLP results indicated that one strain of B. ovis exists in this area. The results showed that the PCR was useful method to investigate the epidemiology of small ruminants' babesiosis. Furthermore, R. Bursa, which can transovarially transmit B. ovis and as well as being widely distributed in West Azerbaijan province, Iran, might play an important role in the field as a natural vector of B. ovis.