Morphological, serological, molecular detection, and phylogenetic analysis of Trypanosoma evansi in horses of different regions in Iran.

Trypanosoma evansi, the causative agent of "surra" is enzootic in Iran. The current study aimed to detect T. evansi in horses from different regions of Iran using morphological, serological, and molecular methods. In 2021, 400 blood samples were collected from horses in eight regions. Eighty horses showed clinical signs such as cachexia (n = 64), fever (n = 36), foot edema (n = 40), and abdominal edema (n = 32), and 320 horses appeared healthy. All samples from the studied regions were evaluated for the presence of trypanosomes using direct analysis of blood smears, mercuric chloride, and PCR-based tests. In total, 12% (95% CI: ± 3.1%), 21% (95% CI: ± 3.9%), and 21% (84) of animals were positive for Trypanosoma in microscopic, serologic, and molecular analyses, respectively. All animals positive for SSU rDNA PCR were from Qom, Semnan, and Golestan regions. Further molecular analyses on 84 PCR-positive horses revealed that 29 horses scored positive in PCR using primers of trypanozoon species and 5 scored positive in PCR using primers of Trypanosoma evansi type A. All samples (n = 5) were from Qom region. The 205-bp fragments of T. evansi RoTat 1.2VSG (accession numbers: ON017789-93) analyzed and compared to other isolates sequence from GenBank BLAST search. It has close similarities with isolates from Pakistan, Egypt, Malaysia, Kenya, and India. Data herein demonstrated that horses from Iran were at high risk of T. evansi infection. Comprehensive control programs, such as those based on the application of repellants and traps, and also, compliance with quarantine standards are recommended for minimizing the risk of the infection.

Neospora caninum, a cause of abortion in donkeys (Equus asinus) in Iran.

Neospora caninum is an Apicomplexan parasite that can cause enormous economic losses due to abortions in cattle. The present study investigated the role of Neospora spp. infection in equine abortion in Iranian donkeys using molecular and phylogenetic analyses. Twenty-nine-aborted fetuses and 29 blood samples from their dams were collected from six different regions in the West and Northwest of Iran. They were tested for N. caninum by PCR at the Nc5 locus, followed by sequencing of five of the PCR products. The overall molecular prevalence was 34.5% in blood samples and the prevalence by DNA detection in the aborted fetuses was 13.8%. Evidence of transplacental transmission from positive jennies to their fetuses was detected in 40% of aborting jennies. Comparison of the five partial Nc5 sequences (227 bp length) exhibited 98-100% similarity with N. caninum GenBank sequences. This is the first molecular study and genetic characterization of N. caninum in Iranian donkeys suggests that N. caninum may be a significant cause of abortion in donkeys.

Assessment of hepatic oxidative damage, paraoxonase-1 activity, and lipid profile in cattle naturally infected with Ba7besia bigemina.

Naturally occurring Babesia bigemina infection in cattle is associated with changes in the status of oxidative stress, trace elements, sialic acid, and cholinesterase activity in blood. However, to date there is no description of hepatic damage in the infected animals. More importantly, the majority of the above-mentioned causative factors are synthesized or stored in the liver. Therefore, this study was undertaken to evaluate biomarkers of hepatic function, paraoxonase-1 activity, and lipid profile in 13 cattle infected with B. bigemina which did not respond to standard treatment. The animals were necropsied and the histopathology of the liver and DNA damage of hepatocytes were examined. Blood analysis revealed a significant parasitemia burden-dependent increase in the activities of hepatic enzymes and total bilirubin and a decrease in albumin concentrations in the infected cattle compared to the control ones. Paraoxonase-1 activity was remarkably lower in the infected animals than the control. A significant decrease in the blood concentrations of total cholesterol, low density lipoprotein, and high density lipoprotein and a significant increase in the triglyceride concentration were observed in the infected animals. Severe oxidative damages were also recorded in the haptic tissue evidenced by significant alterations in the activities of antioxidant enzymes, suppression of total antioxidant capacity, and oxidation of biomolecules. Congestion of blood vessels, bile duct hyperplasia, and hepatocyte necrosis were the evident histopathologic findings. Our results revealed significant changes in the indices of liver function in the diseased cattle, leading to the conclusion that the parasite can potentially cause liver dysfunction.

Evaluation of inflammatory biomarkers in goats naturally infected with Babesia ovis.

This study was designed to evaluate the effects of Babesia ovis infection on concentrations of some essential acute phase proteins (APPs) including albumin, fibrinogen, serum amyloid A, haptoglobin, and ceruloplasmin as well as total, protein-binding, and lipid-binding sialic acids (TSA, PBSA, and LBSA) and two crucial cytokines including interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). Some hematological parameters also were evaluated. Furthermore, any probable correlation among the APPs, SAs, IFN-γ, and TNF-α was calculated. A total of 420 Marghoz and Raeini goats with the ages of 1-3 years old from the north and northwest of Iran were examined, and 17 goats confirmed to be infected with B. ovis by both routine microscopic examination of blood films and molecular assays. As the control, 17 healthy goats were included. The results revealed a significant decrease (P < 0.05) in erythrocyte count, hemoglobin level, and pack cell volume as well as a nonsignificant increase in white blood cell count in the diseased animals compared with the control. Additionally, all the APPs, SAs, and cytokines were remarkably higher in the infected animals than the uninfected ones, except for albumin, which was significantly lower. Moreover, a strong and positive correlation was detected among the parameters mentioned above, except for albumin, which was inversely correlated with the other parameters. In conclusion, B. ovis infection is associated with the induction of severe inflammatory reactions in goats, and both SA and APP are significantly involved in the pathophysiology of the disease.

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.

Serum concentration of sialic acids in naturally occurring ovine babesiosis.

This study is designated to assess the effect of the severity of Babesia ovis infection on sialic acid concentration in blood sera in naturally infected sheep. Infected animals (diseased group) comprised 38 Iranian fat-tailed sheep, about 1-3 years old, naturally infected with B. ovis, divided into four subgroups with respect to parasitemia rates (low 0.1-0.3 %, moderate 0.4-0.9 %, high 1-2.5 %, and very high >2.5 %). The parasitological diagnosis was confirmed using PCR analysis. As a control group, ten clinically healthy sheep reared under the same management and environmental conditions were also sampled. Hematological parameters and the concentrations of total sialic acid (TSA), lipid-bound sialic acid (LBSA), and protein-bound sialic acid (PBSA) were measured in both groups. Compared to controls, sialic acid concentrations showed significant increase (p < 0.05) in infected sheep. Parasitemia rate was positively correlated with sialic acid concentrations. This study demonstrated that B. ovis infection induced marked and persistent elevations of serum sialic acid concentrations. It seems that increase of serum sialic acid concentrations during parasitemia alter receptor-ligand interactions, which are known to play important role in immune response. Furthermore, sialic acid would indirectly inhibit the action of leukocytes and consequently promote the evasion of the immune response and persistence of the parasite in the host. This factor could influence the parasite-host cell adhesion, but further detailed biochemical investigations are needed to precisely explain the exact role of sialic acid in invasion process of the parasite to the host cells.

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.

Molecular detection and phylogenetic analysis of Borrelia spp. from sheep and goats blood samples in West Azerbaijan province, Iran.

Borrelia species are spirochetes transmitted by ticks that are important in human and animals. In most countries, there is still no molecular epidemiology of borreliosis in ruminants. This study was aimed to evaluate the existence of Borrelia spp. DNA in the blood samples of small ruminants using polymerase chain reaction (PCR) method in West Azerbaijan Province, Iran. To detect Borrelia spp. DNA, about 1,018 ruminants (456 goats and 562 sheep) blood samples were examined from different bioclimatic regions in West Azerbaijan province, Iran. The DNA extracting and PCR were conducted. In sheep, the following prevalence rates were respectively obtained for the 16S rRNA, 5S - 23S rRNA and ospA genes: 3.55% (20/562), 2.13% (12/562) and 0.88% (5/562). And so, the prevalence rates of the genes in goats were 0.87% (4/456) for 5S - 23S rRNA gene, 1.75% (8/456) for 16S rRNA gene and 0.65% (3/456) for ospA gene. The prevalence of Borrelia spp. was significantly different in small ruminants based on the farms and localities. The sheep and goats in humid areas (north of West Azerbaijan) were infected statistically more than those in sub-humid areas (south of West Azerbaijan). It is demonstrated that host species like sheep and goats may have a key role in natural Lyme disease cycles and other borreliosis diseases in Iran.

Molecular Identification, Multigenic Characterization and Phylogenetic Analysis of Ctenocephalides Canis Fleas from Humans and Domestic Animals in Iran.

BACKGROUND: Fleas belonging to the Pulicidae are prevalent ectoparasites infesting mammals and birds in Iran. This study focused on genetically identifying and characterizing Ctenocephalides canis collected both off-host and infesting humans and various domestic animals in the country. METHODS: A total of 918 adult flea samples were collected from 10 sites in western and northwestern Iran between April 2018 and May 2019. Out of these, 71 specimens were found off-host, while the remaining fleas were collected from humans (121), sheep (126), goats (184), and dogs (416). Morphological identification at the genus level was performed on all fleas, and ten selected specimens selected based on the sampling sites and hosts were subjected to molecular detection at the species level by using partial amplification and sequencing of the internal transcribed spacer 1 and 2, as well as the cytochrome oxidase I (COXI) markers. RESULTS: The morphological identification confirmed all fleas as Ctenocephalides spp. Alignment and phylogenetic analysis of nuclear and mitochondrial partial sequences confirmed the presence of C. canis. However, molecular divergence was observed among the ten isolates based on the ITS1 and ITS2 with diversity rates estimated at 0.15% and 3.36%, respectively. Notably, the analysis of the COXI marker revealed no molecular divergence among the partial sequences representing the ten studied isolates from C. canis. CONCLUSIONS: This study explores the diversity of C. canis in the western and northwestern regions of Iran, providing insights into their molecular taxonomy and potential role as disease vectors in these areas.

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.

Investigation of pyrethroid resistance mutations in Linognathus stenopsis lice collected from goats in western and northwestern Iran.

Introduction: Linognathus stenopsis lice are an extensive parasitic concern in goat populations worldwide, posing significant economic and health risks. This study examined the identification of alleles of resistance to pyrethroid and mutations in L. stenopsis samples obtained from goats in five provinces in western and northwestern Iran. Methods: Morphological and molecular techniques were employed to identify the louse species. Molecular identification methods and gene sequencing were used to identify resistance-associated mutations in the voltage-gated sodium channel (VGSC) gene. Results and discussion: The results revealed that six amino acid substitutions, including threonine-to-isoleucine (T917I), leucine-to-phenylalanine (L920F), isoleucine-to-phenylalanine (I927F), phenylalanine-to-alanine (F928A), valine-to-arginine (V929R), and arginine-to-leucine (R930L) mutations, were present in the VGSC gene of L. stenopsis lice from various regions of Iran. These findings suggest the potential for pyrethroid resistance development in this louse species, highlighting the importance of integrated pest management (IPM) strategies. Such strategies, which combine selective insecticides, regular grooming, and environmental sanitation, are crucial for effectively managing L. stenopsis infestations and preserving the efficacy of pyrethroids for pest control. Moreover, the emergence of novel kdr mutations underscores the need for ongoing research into the molecular mechanisms underlying these mutations. This research is vital for developing strategies to combat pyrethroid resistance and maintaining the efficacy of insecticides in controlling lice.

Impaired efficacy of fenbendazole and ivermectin against intestinal nematodes in adult horses in Iran.

This study aimed to evaluate the efficacy of fenbendazole and ivermectin on strongyles and Parascaris sp. infecting adult riding horses in three regions with different climates. During 2021 and 2022 fecal specimens were randomly collected from 483 horses older than three years in 31 equestrian clubs in Hamedan (n = 217), Yazd (n = 146) and Tabriz cities (n = 120). Eggs were counted by McMaster technique, and the strongyle larvae were identified using coproculture, PCR and sequencing. Horses with strongyles and Parascaris egg counts ≥150 were enrolled in fecal egg count reduction (FECR) examination following treatment with ivermectin or fenbendazole. In total, 26.5% of examined horses were positive with at least one parasite. Infection rates varied in three cities i.e., 25.8% in Hamedan, 28.8% in Yazd, 25% in Tabriz. Fifty-seven horses had FECR measured. FECR below <90% was observed for IVM-strongyle in two horses in Tabriz, for FBZ-strongyle in two horses in Tabriz and two horses in Hamedan, for IVM-Parascaris in one horse in all three cities, and for FBZ-Parascaris in one horse in Yazd. Furthermore, FECR 90-100% was observed in IVM-Parascaris and FBZ -Parascaris groups in Tabriz. Data herein presented demonstrate different degrees of resistance of strongyles and Parascaris infecting horses in Iran against both ivermectin and fenbendazole. Since non-principled use of anthelminthics is common among horse owners, urgency of test-and-treatment strategy should be educated and implemented by policy-making organizations. Evaluating efficacy of different anthelminthics and choosing the most effective treatment in each region is suggested.

Molecular Detection of Borrelia spp. in Ticks of Sheep and Goats by Nested PCR Method in West Azerbaijan Province, Iran.

Background: Ticks are an important vector among arthropods associated with serious medical and veterinary problems. In this research, we investigate Borrelia species in ticks isolated from the surface of livestock (sheep and goat) in different regions of West Azerbaijan province. Materials and Methods: Polymerase chain reaction (PCR) was performed using specific primers targeting Borrelia spp. genes. Borrelia spp. was identified through PCR. The positive PCR products were sent to Pishgam Company for sequencing. Sequenced data were analyzed, and phylogenetic analysis was performed using maximum likelihood method in MEGA V.10. Results: The detection rate of Borrelia spp. for 16srRNA gene 69 (n = 542; 12.7%; 95%Cl: 10.1%-15.8%), 42 (n = 542; 7.7%; 95%Cl: 5.78%-10.1%) positive on 5S-23SrRNA gene and ospA gene 4 (n = 542; 0.74%; 95%Cl: 0.29%-1.88%). Conclusion: These results are the first report in Iran to identify Borrelia spp. These results of the study showed that the Borrelia spp. in hard ticks detected by PCR and it was negative to soft ticks. it is important in public health implications at the studied areas.

Frequency and genotyping of Giardia duodenalis in dogs of Urmia, northwest of Iran.

Giardia duodenalis is a zoonotic protozoan infecting various vertebrates such as humans and domestic animals. The aim of this study was to determine the frequency and genotypes of G. duodenalis using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in dogs of Urmia, Iran. Overall, 246 stool specimens were collected from 100 pet, 49 stray, and 97 shelter dogs in the Urmia, Iran. Totally, seven samples (2.48%) were microscopically positive in terms of Giardia cyst. The PCR-RFLP analysis revealed that three (1.21%) and two (0.83%) samples have the C and D genotypes, respectively. In addition, two samples (0.83%) were belonged to the AI sub-group. A significant association was determined between the frequency of Giardia infection and life style, age, and stool form of dogs. The findings of the study showed the high frequency of Giardia infection in stray dogs and the dogs under one-year-old. Furthermore, the C and D genotypes of G. duodenalis were predominant in dogs of Urmia, Iran.

Detection of pyrethroids resistance alleles in goat biting louse Bovicola caprae (Phthiraptera: Trichodectidae) in west and northwest of Iran.

Pyrethroid insecticides target voltage-gated sodium channels (VGSCs) that are essential for electrical signaling in the nervous system of insects. Three-point mutations at the corresponding amino acid sequence positions M815I, T917I, and L920F located in domain II conferring the knockdown resistance (kdr) are the most important mutations in pyrethroid-resistant lice worldwide. In addition, six new mutations have been reported in the extracellular loops IIS1-2 (H813P) and IIS5 (I927F, L928A, R929V, L930M, L932M) in the α- subunit of the sodium channel in lice. The aim of this study was to detect alleles resistant to pyrethroids in the domain II (S5-S6) of the VGSC gene in goat biting louse. Goat biting lice were collected from five provinces in the west and northwest of Iran. Genomic DNA was extracted from goat biting lice and Bovicola (Damalinia) caprae species was confirmed by amplifying the cytochrome oxidase subunit I (COXI) gene. A fragment in the domain II (S5-S6) of the VGSC gene was amplified using the specific primers and the resultant polymerase chain reaction products were sequenced. Substitutions T917I, L920F, I927F, L928A, R929V and L930M were identified in the examined sequences. The results showed that all the examined lice had at least one mutation in their VGSC gene associated with pyrethroid resistance or new mutations. The presence of these mutated alleles in the VGSC gene may be due to the long-term and multiple use of pyrethroids against arthropods. Thus, the molecular detection of resistance to pyrethroid insecticides in goat chewing lice can help plot a kdr frequency map to enact effective policies to control caprine pediculosis.

Susceptibility of different life stages of Ornithodoros lahorensis to entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana.

The use of chemicals for the control of arthropod pests can be problematic due to the potential for both environmental contamination and resistance development. As a result, there is an increasing interest in nonchemical alternatives, such as biocontrol by entomopathogenic fungi. In the present study, three strains of Metarhizium anisopliae (V245, 3247, and 4456) and one strain of Beauveria bassiana (LM 3.2) were evaluated under laboratory conditions for their virulence towards three life stages of Ornithodoros lahorensis. Groups of eggs, larvae, and adult ticks were treated by immersion in two different suspensions (10(5) and 10(7) conidia/ml) of each fungal strain. All treatment and control groups were observed during a 3-week period, and the hatchability of eggs and mortality percentage of larvae and adult ticks were assessed. The effect of fungal strains on egg hatchability and larva and adult mortality was significant and dose dependent compared to the control groups (P < 0.05). The results also showed that the greatest biopesticidal effect was due to strain 4456 of M. anisopliae and LM 3.2 strain of B. bassiana at all tested concentrations, making these fungi potential biological control agents of O. lahorensis reducing the use of chemical acaricides.

Pyrethroids resistance in Pulex irritans and Ctenocephalides canis in west and northwest Iran.

Resistance to the knockdown effect of pyrethroid insecticides occurs due to mutations at target sites of pyrethroids, meaning the voltage-gated sodium channels gene (VGSC) in the membrane of the neurons. In fleas, this mutation occurs at two sites in the sodium channel in neurons: one is the replacement of leucine with phenylalanine (L1014F) and the other is the replacement of threonine with valine (T929V). In this study, 81 Pulex irritans and 47 Ctenocephalides canis fleas were collected from five provinces in the west and northwest of Iran. Adult fleas were exposed to cypermethrin 0.75%, and the mortality rate was calculated after 1 and 8 hr, and the mutation sites in the VGSC gene were investigated. The lethality of cypermethrin 0.75% for P. irritans was 40.00 - 57.14% after 1 hr and 60.00 - 73.91% after 8 hr. The lethality of this dose for C. canis after 1 and 8 hr of exposure was 33.33 - 41.17% and 66.66 - 80.33%, respectively. The VGSC sequence analysis indicated two mutation sites in the resistant and one mutation site in the susceptible fleas. The VGSC sequence analysis of susceptible P. irritans showed that 5.50% of them were homozygous susceptible and 94.45% were hetero-zygous susceptible. Susceptible C. canis were 5.26% homozygous and 94.73% heterozygous susceptible. All the resistant fleas were homozygous. The development of pyrethroid resistance and high-frequency L1014F mutation in fleas suggest that pyrethroids are likely to be ineffective in controlling fleas. Therefore, monitoring pyrethroid resistance and its underlying mechanisms are necessary for controlling fleas and finding new alternative control methods.

In vitro assessment of anti-Trichomonas effects of Zingiber officinale and Lavandula angustifolia alcoholic extracts on Trichomonas gallinae.

Trichomonas gallinae is a parasite that acts as a canker-causing agent and leads to significant loss and mortality, especially in young birds. Metronidazole is the approved drug used for the treatment of trichomoniasis. A non-chemical alternativess such as medical plant extracts are also used to treat this disease due to drug resistance. This study aimed to assess in vitro antitrichomonal effects of Lavandula angustifolia and Zingiber officinale extracts on T. gallinae compared with metronidazole. The T. gallinae samples were obtained from infected pigeons. Multi-well plates filled with different concentrations (5.00, 10.00, 25.00, 50.00, and 100 µg mL-1) were used to perform in vitro analysis. The Z. officinale extract's minimum inhibitory concentration (MIC) in the 24-hr period was 25.00 µg mL-1, while it was 50.00 µg mL-1 for metronidazole. The MIC value obtained for L. angustifolia extract in 24-hr was 50.00 µg mL-1. The results indicated that the extracts of Z. officinale and L. angustifolia could act as potential natural agents against trichomoniasis. Furthermore, this study delineated the equal efficiency of L. angustifolia and Z. officinale with that of metronidazole in inhibiting the growth of Trichomonas gallinae trophozoites in culture media.

Babesia canis caused clinical babesiosis in a female Shih Tzu dog.

A 2-year-old female Shih Tzu dog was submitted with the history of anorexia and depression for one week and no prior surgery. Fever and pale mucosa were noticed in physical examination. Microscopic examination of the Giemsa-stained blood smear disclosed large form of Babesia and single to four pear-shaped merozoites within erythrocytes (RBCs). Regenerative anemia characterized by a marked reticulocytosis, significant intra-vascular hemolysis, nucleated RBCs, left-shifted neutrophils, thrombocytopenia, azotemia, high serum creatinine and urea concentrations were recorded following hemato-biochemical analysis. Abundant bilirubin crystals and abnormal reddish color after centrifugation were observed in urinalysis. Molecular analysis was performed using specific primers for detection of Babesia canis. Diminazene aceturate, ciprofloxacin, ivermectin and phosphorus-vitamin B12 were prescribed and the clinical signs improved after four days. Two months follow-up showed no recurrence. Such studies would significantly contribute to the development of appropriate preventive strategies and successful treatment. This communication reports a clinical case of canine babesiosis caused by B. canis in a female Shih Tzu dog.