Molecular identification, risk factor assessment, and phylogenetic analysis of tick-borne pathogens in symptomatic and asymptomatic cattle from South-Eastern Iran.

Tick-borne pathogens (TBPs) represent a substantial threat to cattle globally, exerting adverse impacts on production, health, and economic viability. This study delves into the prevalence and implications of TTBPs in cattle sourced from resource-limited smallholder livestock farms situated in southeastern Iran, proximate to Afghanistan and Pakistan. Blood and tick specimens were systematically collected from a cohort of 230 cattle, comprising 150 asymptomatic and 80 symptomatic individuals. Genomic DNA isolated from blood samples underwent rigorous examination for the presence of key TBPs, including Anaplasma marginale, A. phagocytophilum, A. bovis, A. centrale, Babesia bigemina, and Theileria annulata, utilizing multiple genetic markers. Nucleotide sequence analysis facilitated the reconstruction of phylogenetic relationships. The study also evaluated various potential risk factors, such as clinical status, gender, age, breed, tick infestation, and management practices, to elucidate their associations with TTBPs. Among the cattle cohort, a staggering 87.8% (202/230) tested positive for at least one pathogen. Prevalence statistics encompassed A. marginale (72.2%), T. annulata (68.3%), A. phagocytophilum/A. platys-like complex (66.1%), A. centrale (16.7%), B. bigemina (10.0%), and A. bovis (6.1%). Remarkably, mixed infections involving two, three, and four pathogens were detected in 23%, 52.1%, and 2.2% of animals, respectively. Notably, all asymptomatic cattle were positive for at least one TBP. Tick infestation was observed in 62.2% (143/230) of cattle, predominantly caused by Hyalomma anatolicum (82.5%), Rhipicephalus (Boophilus) annulatus (13.1%), and R. sanguineus sensu lato (4.4%). Risk factors linked to TBPs encompassed tick infestation, older age, and crossbred animals. Clinical presentations among symptomatic cattle encompassed fever, anemia, weight loss, anorexia, jaundice, and enlarged superficial lymph nodes. This study underscores the pivotal role of asymptomatic carriers in the propagation of TTBPs within endemic regions. Furthermore, it emphasizes the potential for the implementation of molecular diagnostics to unmask subclinical infections, thereby affording the opportunity for targeted interventions aimed at ameliorating the burden of TTBPs in resource-constrained smallholder dairy farms.

Molecular detection of Crimean-Congo Haemorrhagic Fever (CCHF) virus in hard ticks from South Khorasan, east of Iran.

BACKGROUND & OBJECTIVES: Crimean-Congo Hemorrhagic Fever (CCHF) is a deadly viral infection reported from more than 30 countries. It is considered a zoonosis↱ and tick bites are the main route of transmission in nature. So far, the virus has been identified in 31 species of hard (Ixodidae) and soft (Argasidae) ticks. The aim of this study was to determine the rate of CCHF virus infection in hard ticks from South-Khorasan province, east of Iran. METHODS: In this study, 684 livestock including 302 sheep, 344 goats, 16 cows and 22 camels were sampled from Birjand, Qaen, Khusf, Darmian and Sarbisheh counties. Genus and species of the ticks were diagnosed under stereomicroscope according to valid morphological keys. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the CCHF virus genome based on S segment in 100 ticks. RESULTS: RT-PCR detected CCHF virus genome in 7 out of 100 ticks. Positive ticks belonged to Hyalomma and Rhipicephalus genera. CCHF virus infected species were Rhipicephalus sanguineus, Hyalomma detritium and Hyalomma asiaticum. All the infected ticks were isolated from goat and sheep and were from Birjand county. INTERPRETATION & CONCLUSION: Our results suggest that Hyalomma and Rhipicephalus may be the main vectors of CCHF virus in the study area.

Influence of ketamine, propofol or isoflurane on intraocular pressure, heart rate and blood pressure in healthy dogs premedicated with medetomidine and midazolam.

BACKGROUND: According to the findings of several studies, sedatives and anaesthetics have different effects on the functioning of the cardiovascular system and intraocular pressure (IOP). For accurate diagnosis, treatment and surgery with minimal complications, it is necessary to be aware of the effects of sedatives and anaesthetics on the cardiovascular system and IOP. OBJECTIVES: The aim of this study was to evaluate the effects of sedatives (medetomidine and midazolam) and anaesthetics (ketamine, propofol and isoflurane) on IOP, heart rate (HR) and blood pressure in dogs. METHODS: In this study, 10 dogs participated in three treatments using a randomised cross-over design, with a 1-week washout period between each treatment. Dogs in all treatments were premedicated with medetomidine and midazolam. Anaesthesia was induced using ketamine, propofol, or isoflurane and maintained for 60 min with the appropriate doses of each drug. The cardiovascular variables (heart rate, and systolic, diastolic and mean arterial pressures) and IOP were measured at different timepoints: before premedication (baseline values, T-Bas), 15 min after medetomidine administration (T-Med), 20 min after midazolam administration (T-Mid) and at 15 (T-15), 30 (T-30), 45 (T-45) and 60 (T-60) min after anaesthesia induction. RESULTS: Medetomidine significantly reduced the IOP and HR and did not significantly change the mean arterial pressure (MAP). Midazolam significantly reduced the IOP while did not significantly change the HR and MAP. Ketamine and isoflurane significantly increased the IOP and HR while did not significantly change the MAP. Propofol significantly increased the HR, but did not cause significant changes in IOP and MAP. CONCLUSIONS: Considering that anaesthetics are typically administered in conjunction with pre-anaesthetic drugs, the increases in IOP induced by ketamine and isoflurane are not important, as the IOP did not exceed the baseline values. However, further studies are required to investigate these effects in patients with elevated IOP.

Exploring the prevalence and impact of Ligula intestinalis infection across fish species in Sistan region, Iran.

Fisheries play a crucial role in aquatic ecosystems, a resource with ecological and economic importance. Measuring parasite prevalence is necessary to assess these fish species' health and well-being. Within the Sistan region, this study sought to estimate the prevalence and intensity of infection with plerocercoid Ligula intestinalis (L. intestinalis) in seven fish species and identify factors such as fish weight, seasonality, and fishing location that might influence infection rates. 2800 fish from seven species were obtained in the Sistan region throughout four seasons. Fish samples from three weight categories (<200gr, 200-400gr, 400gr<) were examined parasitologically. Ligula intestinalis was detected in 96/2800 (3.4%) of fishs. There was a variation in prevalence among fish species. Alburnus charusini (A. charusini), Hypophthalmichthys molitrix (H. molitrix), Hypophthalmichthys nobilis (H. nobilis), and Schizocypris altidorsalis (S. altidorsalis) showed no infection. Schizothorax zarudnyi (S. zarudnyi) showed a prevalence of 1.5% (6/400), Cyprinus carpio (C. carpio) 15% (60/400), and Ctenopharyngodon idella (C. idella) 7.5% (30/400). It was found that infection prevalence was significantly influenced by fish weight in C . carpio and C . idella. A higher prevalence was found in fish with lower weight. Infection rates significantly fluctuated with season across species, but fishing locations had no significant difference. The present study highlights the significant prevalence of L. intestinalis infections in fish populations within Iran's Sistan region, underscoring the need for ongoing monitoring and research to inform effective fishery management strategies. Future studies should focus on long-term monitoring efforts and consider the broader global context of fish parasitology to understand parasite-host interactions better and inform conservation measures worldwide.

Prevalence of Salmonella infection in village chickens and determination of the tetracycline resistance genes in the Salmonella isolates in the Sistan region, Iran.

Recently, an increasing number of multi drug resistant Salmonella species have been emerged due to overuse of antibiotics in veterinary and human medicine which has adverse consequences on public health. The present study was conducted with the aim of investigating the prevalence of Salmonella infection in village chickens in Sistan region and determining the prevalence of the antibiotic resistance genes in Salmonella isolated from these birds. In this study, 100 chickens were randomly selected from five counties of Sistan region. A cloacal swab sample was taken from each bird and also information about age, gender, breed, proximity with other birds, proximity with waterfowl, proximity with livestock, and receiving different antibiotics especially tetracycline were obtained using a questionnaire. Conventional culture methods used for Salmonella detection and isolation. Then, amplification of invA gene by PCR was used to confirm Salmonella colonies. Finally, 27 samples were confirmed to be infected with Salmonella by both culture and PCR methods. Disk diffusion method was used to determine the sensitivity to 4 antibiotics including; tetracycline, gentamicin, cefepime, and difloxacin. The results of the present study showed that proximity to waterfowl (OR = 0.273) significantly mitigates the risk of Salmonella infection. For the isolates, the highest resistance was recorded against cefepime and the highest susceptibility was to difloxacin. The presence proportion of tetA and tetB in tetracycline resistant isolates was higher than that in susceptible ones but this difference was not statistically significant.

Effects of dietary vinegar on performance, immune response and small intestine histomorphology in 1- to 28-day broiler chickens.

The purpose of this research was to evaluate the effects of apple cider vinegar in diet on the growth performance, immune response, histomorphological changes of the small intestine and some serum biochemical factors in broilers. A total of 85 broiler chicks of Ross 308 were purchased and 64 well-conditioned chicks were selected and divided into four experimental groups with four replicates and four chicks per replicate. The diets of groups 2, 3 and 4 were mixed with 1, 2 and 3% of vinegar, respectively, and group 1 as the control group was fed by the standard diet. The results showed that body weight gain was higher in the groups fed vinegar than the control group. There were no significant changes in the feed conversion ratio between the treatment and control groups (p = .507). Vinegar intake through the diet did not change significantly the weight of Bursa of Fabricius (p = .369) and spleen (p = .122). Vinegar significantly reduced blood urea nitrogen levels in the treatment groups compared with the control group (p = .0052). There was a significant increase in the level of antibody titre against Newcastle disease virus by haemagglutination inhibition test in the groups receiving vinegar in comparison with control group (p = .0358). Compared with the control group, the villus height (p = .0022) and intestinal crypts depth (p = .0015) significantly increased in the groups receiving apple cider vinegar. In conclusion, dietary supplementation with apple cider vinegar has beneficial effects on performance, immune response and small intestine histomorphology in broilers.

Identification of Anaplasma marginale in long-eared hedgehogs (Hemiechinus auritus) and their Rhipicephalus turanicus ticks in Iran.

Hedgehogs play a role in the eco-epidemiology of parasites, viruses and bacteria of veterinary and medical importance. In Europe, hedgehogs have been found infected with tick-transmitted Anaplasma phagocytophilum and A. marginale. In this study blood and ticks collected from 53 long-eared hedgehogs (Hemiechinus auritus) living in southeastern Iran close to Afghanistan and Pakistan borders were examined for Anaplasma spp. infection using microscopical and molecular biology methods. At microscopical examination Anaplasma-like inclusion bodies were found at the margin of erythrocytes in the blood smear of one infected hedgehog (prevalence 1.9 %). Each hedgehog was infested with three to eight adult ticks (average 4.7 ± 1.5). On the body of hedgehogs, the ears were the main site for attachment of ticks. Out of 248 collected adult ticks 81.4 % were Rhipicephalus sanguineus sensu lato and 18.5 % were R. turanicus. PCR and sequencing showed the presence of A. marginale infection in the blood of two hedgehogs (3.8 %) and R. turanicus ticks collected from them. This is the first report of A. marginale infection in hedgehogs in Iran. Epidemiological importance of this finding is discussed and current knowledge on the tick fauna of hedgehogs in the country is reviewed.

Prevalence of dirofilariasis in shepherd and stray dogs in Iranshahr, southeast of Iran.

Dirofilariasis is a zoonotic parasitic disease, which its cause Dirofilaria immitis, a nematode transmitted by insects and a worldwide dissemination. Dogs and cats are the main hosts of this parasite. Therefore, this study was conducted to determine the prevalence of D. immitis in shepherd and stray dogs in Iranshahr city, southeast of Iran. In this study 49 shepherd dogs and 50 stray dogs selected randomly and the blood samples were taken and sent to parasitology laboratory. Thin and thick thin blood samples were prepared and stained with Giemsa method and modified knott method. The obtained data were analyzed by SPSS 18 statistical software. The overall prevalence of parasite was 30.3%, seven of 49 shepherd dogs (14/3%) and twenty three of 50 stray dogs (46%) were positive. From 30 positive samples, 14.3% of shepherd dogs and 50% of stray dogs were male, and 44.7% of stray dogs were female. 14% of infected shepherd dogs and 38.5% of infected stray dogs were indigenous, and 16.7% of infected shepherd dogs and 72.7% of infected stray dogs were hybrids. There was no statistically significant relationship between the prevalence of D. immitis with age, gender, breed and usage of anti-parasitic drugs. Due to the relatively high contamination of dogs in the city of Iranshahr with this parasite, veterinarians and public health professionals should be considered necessary and preventive measures in relation to this disease.

Epidemiology and Molecular Prevalence of Toxoplasma gondii in Cattle Slaughtered in Zahedan and Zabol Districts, South East of Iran.

BACKGROUND: Toxoplasma gondii is an obligate, intracellular parasite which causes the toxoplasmosis in humans and warm-blooded animals. Red meat is an important source for transmission of the infection to humans. This study aimed to determine the prevalence of Toxoplasma among imported and indigenous cattle in the Sistan region. METHODS: One hundred samples from slaughtered cattle were collected from two abattoirs of Zabol and Zahedan, South East of Iran in 2015. Each sample was a mixture of three muscle, including tongue, cardiac, and triceps. Additional data of each cattle, including sex, breed, age, indigenous or imported, location of slaughter, management practices, and feeding system were obtained through observations and interviews. Infection by T. gondii was determined by PCR method. RESULTS: The prevalence of Toxoplasma in indigenous cattle was 6% and in imported cattle was 26%, and this difference was statistically significant (P=0.006). Moreover, the prevalence of Toxoplasma was statistically associated with management practices (P=0.01) and feeding system (P=0.001). However, relationship between the prevalence of Toxoplasma with age, breed, sex, and location of slaughter was not statistically significant. CONCLUSION: Since the prevalence of Toxoplasma among imported cattle is higher than indigenous cattle, so strict supervision for importing livestock from neighboring countries is necessary.

Detection of Theileria annulata carriers in Holstein-Friesian (Bos taurus taurus) and Sistani (Bos taurus indicus) cattle breeds by polymerase chain reaction in Sistan region, Iran.

Theileria annulata is common in tropical and subtropical regions especially in Iran and causes great economic losses in cattle industry. In Iran the epidemiological aspects of bovine theileriosis in different breeds of cattle is poorly understood. The aim of present study is comparison of the number of T. annulata carriers in the two major cattle breeds (Holstein-Friesian and Sistani) in Sistan of Iran by giemsa and polymerase chain reaction (PCR) methods. During winter 2013, 160 native cattle, from the two major breeds in Sistan, with the mean age of more than one year and without typical clinical symptoms of theileriosis were selected. At first, a thin layer smear was held from their ear sublime vein blood for Giemsa staining method. In order to do PCR assay, jugular vein blood sample of each cow was taken. The PCR employs primers specific for the 721-bp gene fragment encoding the 30-kDa major merozoite surface antigen of T. annulata. By PCR method, 38 (47.5 %) Holstein blood samples and 22 (27.5 %) Sistani blood samples had DNA of T. annulata and considered positive (The correlation was significant at values of P < 0.05). By checking 160 blood smears with light microscope and lens × 100, only 10 samples (6.25 %) were positive for T. annulata. Statistical comparison between PCR and smear method showed that the PCR method is more sensitive and accurate in comparison to Giemsa staining method to diagnose the asymptomatic carriers of T. annulata.

A study on prevalence and identification of Ovine Theileria and Babesia infection in Zabol using PCR method.

Infection with Babesia and Theileria Causes high mortality and economical losses in livestock and has a relatively high prevalence in Iran. In Zabol, animals are susceptible to this disease because of presence of vector ticks, weather conditions and smuggle animal across the border and they cause great damages to the economy and production. However, few studies have been done for differentiation of Theileria and Babesia in sheep in this area. The purpose of the present study was to determine the prevalence of Babesiosis and Theileriosis in sheep in Zabol. A number of 80 animals were randomly selected and their blood samples were examined. The presence of Theileria and Babesia parasites in each sample was determined with PCR and microscopic examination. Of 80 blood samples obtained from sheep, 8 cases (10 %) were positive by microscopic examination where 6 samples were infected with Theileria and 2 samples with Babesia. The result of PCR method determined the prevalence of 66.25 % (53 samples) and 3.75 % (3 samples) for Theileria and Babesia, respectively. The correlation between these two methods (PCR and microscopic examination) was determined with Kappa statistical test. Based on the obtained results, it is concluded that Babesiosis has a low prevalence among the sheep of Zabol. This is the first report in which ovine Babesiosis has been studied in this region using molecular identification techniques.

A survey on hydatidosis in livestock in Northern Iran based on data collected from slaughterhouses from 2004 to 2008.

Data have been collected from slaughterhouses in three provinces across the Northern Iran (Gilan, Mazandaran and Golestan) from March 2004 to March 2008. These data were collected to evaluate the prevalence of hydatidosis in slaughtered cattle, sheep and goats. During the study, 3,347,797 animals were slaughtered. These included 621,686 cattle, 1,719,725 sheep and 1,006,386 goats. The prevalence of infection in cattle, sheep and goats was 12%, 14.6% and 10.1%, respectively. The association of condemnation rates with seasons was not proven statistically.