Babesia Ovis Research Articles

Assessment of Babesia ovis pathogenicity in goats: implications for transmission dynamics and host resistant

Babesia ovis, commonly associated with ovine babesiosis, poses a significant threat to sheep health, often resulting in severe clinical manifestations and high mortality rates. However, the impact of B. ovis on goats has remained uncertain, prompting us to investigate its pathogenicity in caprine hosts. Experimental infections using B. ovis-infected blood inoculation and infected tick infestation, were conducted on spleen-intact (n=5) and splenectomized (n=5) goats. The experimental infection was performed using fresh blood obtained from a B. ovis-infected splenectomized sheep. One spleen-intact sheep served as a control for the experimental infection with B. ovis-infected Rhipicephalus bursa ticks. While all experimentally infected sheep (#501, #575) displayed severe clinical symptoms and high parasitemia, goats exhibited resistance, showing no significant clinical manifestations or sustained parasitemia. Notably, B. ovis was detected in two spleen-intact goats via nested PCR, prompting further investigation into their role as reservoirs for tick-borne transmission. These goats were then infested with Babesia spp.-free R. bursa larvae (0.1 gr) and adults (50 females and 50 males) for transstadial and transovarial transmission experiments respectively. Results indicated that chronically B. ovis-infected spleen-intact goats are not significant sources for maintaining the tick-borne transmission cycle of the parasite. These findings highlight the differential susceptibility of goats to B. ovis infection compared to sheep and their limited role as reservoirs for parasite transmission. Understanding the role of goats in B. ovis transmission and their resistance mechanisms can inform effective control measures and reduce economic losses in affected regions. Further research into caprine babesiosis and host immunological responses is essential to fully elucidate their possible role as reservoirs of the parasite, and underlying mechanisms of host susceptibility and parasite pathogenesis.

Mitochondrial Genome Analysis of Babesia ovis (Apicomplexa: Babesiidae) Endemic in Sheep in Türkiye

This study presents the first comprehensive analysis of the mitochondrial genome of Babesia ovis, a significant pathogen in ovine babesiosis in Türkiye. The B. ovis mitochondrial genome is a linear monomeric molecule of 6015 bp with an A + T content of 70.5%, featuring terminal inverted repeats (TIRs) at both ends. It encodes three essential proteins (Cox1, Cox3, and Cob) and six fragments of large subunit rRNA genes. Comparative analysis revealed high sequence identity with Babesia sp. Xinjiang (87.5%) and Babesia sp. Dunhuang (87.5%), suggesting a close evolutionary relationship. The study highlights the conservation of mitochondrial gene content across Babesia and Theileria species, emphasizing their utility in phylogenetic studies. The findings also suggest that TIR variability plays a role in mitochondrial genome size differences, influencing species-specific adaptations. This research provides valuable insights into the evolution and functional adaptations of B. ovis and underscores the potential of mitochondrial genomic data in enhancing diagnostic and therapeutic strategies for babesiosis. Further exploration of mitochondrial genomes in Babesia species is essential for understanding their biology and developing effective control measures.

Detection of Babesia spp., and Theileria spp., in sheep across diverse provinces of Iran

This study aimed to identify the diversity of Babesia and Theileria species in sheep across various regions of Iran using microscopic and molecular techniques, including species-specific PCR and enzymatic digestion. A total of 373 blood samples were collected from sheep during the tick vector activity period, from 2018 to 2021, in provinces such as Tehran, Alborz, Qazvin, Hamedan, West Azerbaijan, Kerman, and Fars.Results showed that 101 samples (27 %) exhibited piroplasms including 78 samples of Theileria spp. and 23 samples of Babesia spp. A molecular approach using general primers detected piroplasm parasites in 145 samples (38 %). Theileria ovis was notably present in 91 samples (24.39 %), followed by Theileria lestoquardi in 24 samples (6.43 %). Babesia ovis infection was detected in 30 samples (8.4 %). Despite extensive molecular evaluation, no other Babesia species, including Babesia motasi, were identified. Co-infections involving T. ovis and T. lestoquardi (4 samples; 1.07 %) and T. ovis and B. ovis (6 samples; 1.60 %) were observed. No Babesia spp. were detected in Kerman and Fars provinces, although T. ovis and T. lestoquardi were present.Blast analysis of the sequences indicated 100 % intra-species similarity, with inter-species similarities of 83.3 % (B. ovis and T. lestoquardi), 84.4 % (B. ovis and T. ovis), and 96.44 % (T. ovis and T. lestoquardi). In conclusion, B. ovis was the main cause of Babesiosis, while Theileriosis was predominantly attributed to T. ovis and T. lestoquardi. Molecular diagnostics play a key role in accurately distinguishing between these species, particularly in cases of co-infection involving Babesia spp. and Theileria spp.

Investigation of Babesia species using molecular methods

Aims: To determine the presence of Babesia species in ticks in the Ankara region and neighboring provinces; the aim of this study was to detect and distribute Babesia species using molecular methods in ticks collected from nature and hosts. Methods: Babesia was detected at the species level by conventional PCR method by DNA extraction of 191 tick samples collected from Ankara and neighboring provinces, which applied to the Parasitology National Reference Laboratory of the Ministry of Health, General Directorate of Public Health, between 2020-2021. Results: A total of 191 tick samples were examined in the study, and as a result of the conventional PCR study, 4 Babesia agents were found positive, 1 of them was Babesia bigemina (B. bigemina), 2 were Babesia ovis, 1 was Babesia spp. were determined. Conclusion: In tick-borne diseases such as Babesiosis; it should not be forgotten that ticks continue to focus some infections in nature and serve as reservoirs for disease agents in nature, and the necessary strategies for tick control should be followed.

High genetic diversity of Anaplasma ovis in sheep from Bosnia and Herzegovina.

Ovine anaplasmosis (sensu stricto) is a rickettsial blood disease caused by the tick-borne species Anaplasma ovis. The disease is characterized by mild anemia, fever, and icterus. A more severe clinical presentation is possible in non-endemic areas. There is no existing data on the presence of Anaplasma ovis in Bosnia and Herzegovina. However, given the country's location within the Mediterranean Basin and the recent molecular detection of Babesia ovis, it is plausible that sheep in the region could naturally be infected with this tick-borne pathogen. Blood samples from 81 sheep in the Podrinje and Herzegovina areas were examined by PCR. PCR positivity was found in 38 (46.9%) cases indicating a high number of infected sheep. Mixed infections with Babesia ovis and A.ovis were observed in 63.3% of cases. A higher number of positive sheep was recorded in the area of Herzegovina. Phylogenetic analysis of the gltA, groEL, and msp4 genes of A. ovis revealed numerous genotypes and significant genetic variability. This diversity was not related to geographic origin, tick-borne infection status, or sheep breeding practices in Podrinje and Herzegovina. The data obtained in this study suggest that the emergence of new genotypes and the high genetic variability of A. ovis are driven by specific local and micro-environmental factors.

Feline vector-borne haemopathogens in Türkiye: the first molecular detection of Mycoplasma wenyonii and ongoing Babesia ovis DNA presence in unspecific hosts

BackgroundCats are hosts and reservoirs for many haemopathogens such as piroplasms, Rickettsia, hemotropic Mycoplasma, Bartonella, Ehrlichia, and Anaplasma, which are transmitted by various vector arthropods and some of which have a zoonotic concern. Although it is noteworthy that the rate of ownership of companion animals has increased in Türkiye in recent years and that cats account for a large proportion of these animals, there is limited research on the vector-borne infectious agents carried by them. The present study aimed to provide a comprehensive molecular epidemiological data and molecular characterization of feline vector-borne haemopathogens (FVBHs), including piroplasms, anaplasmataceae, rickettsias, haemoplasmas, and Bartonella species in Türkiye. In total, 250 feline blood samples were collected from client-owned cats (n = 203) and shelter cats (n = 47) brought to the Small Animal Hospital of Selcuk University, Veterinary Faculty.ResultsOverall, 40 (16%) cats were found to be infected with at least one of the investigated haemopathogens and piroplasm, Mycoplasma spp. and Bartonella spp. prevalence was 1.6%, 11.2%, and 4.8%, respectively. No Anaplasma/Ehrlichia spp. and Rickettsia spp. DNA was detected in the investigated feline samples. Sequence analysis revealed that all four piroplasms belonged to Babesia ovis with a 97.93–99.82% nucleotide sequence identity to 18S rRNA gene sequences from Spain and Türkiye, while some sequenced hemoplasmas were Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm) and Mycoplasma wenyonii, and Bartonella spp. were Bartonella henselae and Bartonella koehlerae species. Co-infections with Mycoplasma spp. and Bartonella spp. were also detected in 4 cats (1.6%) in this study, where single infections were predominant.ConclusionThis study provides valuable information on zoonotically important feline vector-borne hemopathogens in Türkiye, some of which have received attention under the One Health perspective, and is the first molecular epidemiological study to demonstrate the presence of Babesia ovis, the causative agent of ovine babesiosis, and Mycoplasma wenyonii DNA, the causative agent of bovine haemotropic mycoplasmosis, in cats. Further studies on the roles of such pathogens detected in unspecific hosts and the host specificity of the vectors that transmit them will contribute to the elucidation of this situation.

Molecular prevalence and distribution of tick-borne bacterial and protozoan pathogens of sheep and goats in Africa: A systematic review and meta-analysis

Tick-borne pathogens (TBPs) are a major impediment to the health, welfare, and production of small ruminants across the world, including Africa. Comprehensive information about the epidemiology of TBPs infecting sheep and goats across Africa is lacking. Therefore, this study was undertaken to determine the prevalence through a meta-analysis of selected TBPs in blood DNA from domestic sheep and goats in Africa obtained using molecular-based methods. The literature review was done according to Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines using five English electronic databases (PubMed, Science Direct, Springer Link, Web of Science, and African Journals Online (AJOL). The search was performed with no restriction in time through to 18th January 2023. Of the 63 full-text articles subjected to eligibility, only 30 articles met the eligibility criteria and were included in the review. The overall pooled prevalence of selected TBPs varied considerably between host species (sheep vs. goats), with Anaplasma ovis (44.50 vs. 48.40%), Ehrlichia ruminantium (5.50 vs. 2.00%), Coxiella burnetii (4.40 vs. 1.70%), Borrelia theileri in sheep (5.20%), Babesia ovis (1.70% vs. 1.90%), Theileria ovis (40.50% vs 10.00%), T. separata (1.00% vs 1.00%) and T. lestoquardi in sheep (8.40%). However, the prevalence of the selected TBPs was generally higher in sheep compared to goats. Several genetic loci were targeted in the characterization of tick-borne pathogens, such as 16S rDNA, groEL, and msp4 for Anaplasma ovis, pCS20 for Ehrlichia ruminantium, Insertion Sequence (IS1111) for Coxiella burnetii, flaB (flagellin) and 16S rRNA for Borrelia theileri, 5.8S rRNA and 18S rRNA for Babesia/Theileria, as well as the utilization of numerous PCR variants including conventional polymerase chain reaction (PCR), nested-PCR, qPCR, Loop-mediated isothermal amplification (LAMP), and reverse line blotting (RLB). In conclusion, A. ovis was the most widely distributed and prevalent TBP affecting small ruminants within the continent. Hence, this warrants adequate attention towards early diagnosis and treatment of infected animals as well as the control of the tick vectors involved in their transmission.

Seroprevalence and Endemic Status of Babesia ovis by Enzyme-Linked Immunosorbent Assay in East Azerbaijan Province, North-West of Iran.

Babesia ovis, an intraerythrocytic parasite carried by ticks and one of the most common subclinical ovine illnesses, was studied to ascertain its seroprevalence and endemic status in ram and ewe populations in East Azerbaijan Province, Iran, in lambs, yearlings, and adults of over two years of age. A total of 960 sheep from 10 cities were selected from Jan 2018 to Nov 2019. Blood samples were collected from each animal and tested for the presence of B. ovis antibodies by applying a developed enzyme-linked immunosorbent assay (ELISA) technique. Checkerboard titrations were used to determine the optimal dilution of the antigen using negative and positive control sera. To determine whether the disease is endemically stable, inoculation rates for each age group were also calculated. Correlation coefficients were calculated between age and infection rates and also between age and inoculation rates. The results revealed an average infection rate of 49.4% in East Azerbaijan Province. There was a positive correlation between the age of animals and susceptibility to infection except for lambs and yearlings, whereas there was no meaningful difference in exposure to B. ovis between rams and ewes. The negative correlation between age and inoculation rates indicates increased disease instability with age. Inoculation rate results revealed the endemically instable status of B. ovis in the studied area. High prevalence rates and endemically instable status of the disease suggest demand for vaccine development and implementation of appropriate control measures for ovine babesiosis to mitigate the associated economic losses.

Evaluating serum electrolyte and trace element variations between Babesia ovis-infected and uninfected Lohi sheep

Babesiosis caused by Babesia ovis is a major threat to the livestock industry worldwide. In Pakistan, the Lohi sheep breed is an important economic resource, but limited information is available on the impact of B. ovis infection on this breed. This study aimed to investigate the serum electrolyte and trace element variations between B. ovis-infected and uninfected Lohi sheep from Pakistan. A total of 97 Lohi sheep was stratified based on the geographical distribution of Multan district, employing a multistage cluster sampling method. Blood and serum samples were collected from randomly selected sheep, and DNA extraction and PCR amplification were performed to detect B. ovis infection. Serum electrolyte and trace element levels were analyzed in infected (n=67) and uninfected (n=30) groups, stratified by age and gender. The study revealed a 69.07% overall infection rate of B. ovis in Lohi sheep. Infected sheep showed significantly elevated serum copper levels (p<0.0001), with no substantial differences observed in serum iron, sodium, potassium, and chloride levels. However, age-wise variance analysis revealed statistically significant variations in sodium and potassium levels (p<0.0001 and p<0.05, respectively). Notably, serum chloride levels differed between infected and uninfected females in individuals younger than two years (p<0.05). Serum iron levels remained consistent across different age groups. Comparative analysis indicated a higher prevalence of electrolyte imbalances, such as hyponatremia, hyperkalemia, hyperchloremia, and hypoferremia, in infected sheep compared to normal reference ranges, while instances of hypernatremia, hypokalemia, hypochloremia, and hyperferremia were less frequent. In conclusion, our study suggests that B. ovis infections could lead to alterations in serum electrolyte and trace element levels in Lohi sheep, emphasizing the importance of further research into the specific mechanisms driving these alterations to enhance disease management strategies tailored to this breed.

Epidemiological and Molecular Characteristics of Piroplasmids and Anaplasma spp. in Tan Sheep, Ningxia, Northwest China

Piroplasmosis and anaplasmosis are important zoonotic diseases of animal origin, which can be transmitted by ticks to infect animals. However, there is limited information on the infection of piroplasmosis and anaplasmosis in Tan sheep in Ningxia, China. In order to understand the prevalence of piroplasmosis and anaplasmosis in Tan sheep in Ningxia, 150 blood samples of Tan sheep from farms in five urban areas of Ningxia were detected by PCR, and some positive samples were sequenced to establish a phylogenetic tree. PCR revealed that the prevalence of Anaplasma spp. in Tan sheep in Ningxia was 28.0%. The overall prevalence of Piroplasmids was 33.3%, of which Theileria spp. and Babesia spp. were 20.7% and 12.7%, respectively. Among the samples of different ages, the highest detection rates of Piroplasmids and Anaplasma spp. were found in Tan sheep aged 20–30 months, and the detection rate of Theileria spp., Babesia spp., and Anaplasma spp. were 25.4%, 23.6%, and 36.3%, respectively. In this study, one Theileria species was identified as Theileria uilenbergi, two Babesia species were identified as Babesia molasi and Babesia ovis, and two Anaplasma species were identified as Anaplasma ovis and Anaplasma phagocytophilum, and the dominant species were A. ovis and T. uilenbergi. To the best of our knowledge, this is the first report detailing the infection rate and genotype of Piroplasmids and Anaplasma spp. in Tan sheep in Ningxia, China. The results of this study provide valuable data for understanding the epidemiology of tick-borne disease in Tan sheep in Ningxia, China, and lay a theoretical foundation for the prevention and control of piroplasmosis and anaplasmosis in Tan sheep in Ningxia, northwest China.

Serological and Molecular Survey of Babesia ovis in Healthy Sheep in Türkiye

Babesiosis, caused by Babesia ovis, is a major seasonal issue in sheep, particularly in countries like Türkiye with high Rhipicephalus bursa tick populations. Previous studies employing various methods such as microscopy, serology, or molecular techniques have reported different epidemiological data concerning ovine babesiosis. Addressing this knowledge gap, our study employed a combined nested PCR (nPCR)/indirect ELISA (iELISA) approach, analyzing blood samples collected from 414 sheep between April and July 2023 using both techniques. nPCR amplified the 18S ribosomal RNA gene of B. ovis and determined a molecular prevalence of 1.9%. Conversely, serological testing using iELISA targeted the BoSA1 antigen and revealed a significantly higher positivity rate of 59.9% for anti-B. ovis antibodies. The temporary presence of Babesia after recovery reduces nPCR sensitivity, resulting in lower molecular prevalence. However, even if Babesia is not present in the host, anti-B. ovis antibodies remain in the serum for a long time and can be detected serologically. Our study underscores the necessity of concurrently employing molecular and serological methods for an accurate assessment of B. ovis prevalence. It highlights the importance of comprehensive epidemiological approaches for effective disease management in sheep populations.

First molecular survey of tick-borne protozoan and bacterial pathogens in the questing tick population in Bangladesh

Questing ticks carry various tick-borne pathogens (TBPs) that are responsible for causing tick-borne diseases (TBDs) in humans and animals around the globe, especially in the tropics and sub-tropics. Information on the distribution of ticks and TBPs in a specific geography is crucial for the formulation of mitigation measures against TBDs. Therefore, this study aimed to survey the TBPs in the questing tick population in Bangladesh. A total of 2748 questing hard ticks were collected from the pastures in Sylhet, Bandarban, Sirajganj, Dhaka, and Mymensingh districts through the flagging method. After morphological identification, the ticks were grouped into 142 pools based on their species, sexes, life stages, and collection sites. The genomic DNA extracted from tick specimens was screened for 14 pathogens, namely Babesia bigemina (AMA-1), Babesia bovis (RAP-1), Babesia naoakii (AMA-1), Babesia ovis (18S rRNA), Theileria luwenshuni (18S rRNA), Theileria annulata (Tams-1), Theileria orientalis (MPSP), Anaplasma marginale (groEL), Anaplasma phagocytophilum (16S rRNA), Anaplasma bovis (16S rRNA), Anaplasma platys (16S rRNA), Ehrlichia spp. (16S rRNA), Rickettsia spp. (gltA), and Borrelia (Bo.) spp. (flagellin B) using genus and species-specific polymerase chain reaction (PCR) assays. The prevalence of the detected pathogens was calculated using the maximum likelihood method (MLE) with 95 % confidence interval (CI). Among 2748 ixodid ticks, 2332 (84.86 %) and 416 (15.14 %) were identified as Haemaphysalis bispinosa and Rhipicephalus microplus, respectively. Haemaphysalis bispinosa was found to carry all the seven detected pathogens, while larvae of R. microplus were found to carry only Bo. theileri. Among the TBPs, the highest detection rate was observed in A. bovis (20/142 pools, 0.81 %, CI: 0.51–1.20), followed by T. orientalis (19/142 pools, 0.72 %, CI: 0.44–1.09), T. luwenshuni (9/142 pools, 0.34 %, CI: 0.16–0.62), B. ovis (4/142 pools, 0.15 %, CI: 0.05 – 0.34) and Bo. theileri (4/142 pools, 0.15 %, CI: 0.05–0.34), Ehrlichia ewingii (3/142 pools, 0.11 %, CI: 0.03–0.29), and Babesia bigemina (1/142, 0.04 %, CI: 0.00 – 0.16). This study reports the existence of T. luwenshuni, E. ewingii, and Bo. theileri in Bangladesh for the first time. The novel findings of this study are the foremost documentation of transovarian transmission of B. bigemina and E. ewingii in H. bispinosa and also provide primary molecular evidence on the presence of E. ewingii and Bo. theileri in H. bispinosa. Therefore, this study may shed light on the circulating TBPs in ticks in the natural environment and thereby advocate awareness among physicians and veterinarians to control and prevent TBDs in Bangladesh.

The Taxonomıc Research of the Prımıtıve Blood Parasıtes and Transmıttıng Ixodıdae Tıcks of the Small Ruminants in the Shırvan-Salyan Economıc Regıon of Azerbaıjan

The primitive blood parasites - piroplasmids are pathogens transmitted to the agricultural animals by the ticks and caused the death and serious economic damage. In Azerbaijan, in the economic regions where the animal husbandry is developed, the systematic investigation and epizootology of these parasites in small ruminants, were researched by us for the first time. For this purpose, the research work was conducted in the Shirvan-Salyan economic region. For this purpose, researches were conducted on the taxonomic study of primitive blood parasites and their transmitters - Ixodidae ticks of small ruminants in the Shirvan-Salyan economic region. Out of 887 sick and suspected diseases sheep, 483 heads (54.5%), and 125 heads (27.4%) of 456 goats had positive results of peripheral blood smear samples attributed to primitive blood parasites. Babesia ovis and Anaplasma ovis parasites were detected in the erythrocytes of sick sheep, and the infection was mostly associated. A.ovis parasite was detected in sero samples of goats. The organs of 56 sheep and 31 goats that died of high fever - spleen, kidney, liver - were examined for primitive blood parasites. The results of the classical examination method in the diagnosis of A.ovis and B.ovis parasites were compared with the effectiveness of the polymerase chain reaction (PCR). 2,875 sheep and goats (1,786 sheep, 1,089 goats) were examined for ectoparasites in livestock farms, 63.8 percent of sheep and 53.6 percent of goats were intensively infected with ticks. Tick infestation was mostly covered in spring, summer and autumn with high intensity. Rhipicephalus bursa ticks were dominant in sheep and Hyalomma plumbeum ticks in goats. In order to determine transovarial and transstadial transmission of disease agents, smears made from internal organs (salivary gland, ovary, intestine) and eggs of ticks were studied for A.ovis and B.ovis parasites.

Role of Rhipicephalus bursa larvae in transstadial transmission and endemicity of Babesia ovis in chronically infected sheep.

Babesia ovis, transmitted by Rhipicephalus bursa ticks, is the causative agent of ovine babesiosis, a disease characterized by fever, anemia, hemoglobinuria, and high mortality in sheep. This study investigates whether sheep that survived babesiosis without treatment can serve as a source of infection for B. ovis-free host-seeking R. bursa larvae in a later season. Three donor sheep were experimentally infected with B. ovis, and after six months, persistence of B. ovis was assessed through blood and tick transmission experiments. Blood from donor sheep was intravenously injected into three recipient sheep, while donor sheep were also infested with B. ovis-free R. bursa larvae. Engorged nymphs molted to adults, and new recipient sheep were infested with these ticks. All recipient sheep were monitored for B. ovis for 100 days using microscopic, serological, and molecular approaches. The presence of B. ovis was confirmed in the recipient sheep that received blood, leading to clinical infection in two. However, no B. ovis was detected in recipient sheep infested with ticks. These results suggest that sheep recovering from B. ovis infection do not serve as a source of infection for R. bursa larvae in subsequent seasons.

Microscopic and molecular detection of piroplasms among sheep in Upper Egypt.

Blood parasites pose a significant threat to livestock production in southern Egypt, yet there is a scarcity of information regarding their circulation and epidemiology in sheep in this region. This study aimed to investigate the seroprevalence of blood parasite infections in sheep in Assiut governorate, Upper Egypt. A total of 400 blood samples were collected from sheep of varying ages and genders. The preliminary screening for the presence of piroplasms, mainly Babesia and Theileria spp., via microscopic examination, followed by investigation of the potential risk factors linked with the exposure to infection. Moreover, molecular identification of both parasites on some of positive samples was performed using PCR targeting Babesia 18S rRNA and Theileria annulata Tams1 gene. The microscopic examination revealed that among the examined sheep, there was an overall prevalence of blood parasites at 44% (176 out of 400), with Babesia spp. observed in 14% (56 out of 400) and Theileria spp. in 30% (120 out of 400). Furthermore, the infection rate was non-significantly higher in young animals (50%) compared to adults (38.5%) (P = 0.246). Male sheep exhibited a significantly higher vulnerability to both parasites' infection (63.3%) compared to females (35.7%) (P = 0.011). Interestingly, the prevalence of both blood parasites was significantly higher during the cold season (66.1%) compared to the hot season (15.9%) (P = < 0.001). The molecular analysis identified the presence of Babesia ovis and Theileria annulata among a subsample of the positive sheep's bloods films. The identified species were recorded in the GenBank™ databases and assigned specific accession numbers (OQ360720 and OQ360719 for B. ovis), and (OP991838 for T. annulata). Taken together, this study confirms a high prevalence of piroplasmosis and offers epidemiological and molecular insights into blood parasites in sheep from Upper Egypt, highlighting the importance of detecting these parasites in various hosts and their competent vectors (ticks).

The Molecular Detection of the Anaplasma ovis Pathogens of the Serological Samples in Small Ruminants and Ixodid Ticks in Azerbaijan

A total of 704 blood samples (561 sheep, 143 goats) were examined for the Anaplasma ovis infection during a 1-year period. PCR and ELISA were used for the detection of the A. ovis antibodies. PCR assay identified A. ovis in 20 (8.1%) sheep and 15 (6.09%) goats. Using ELISA assay, 8.53% (31) were positive (21 sheep, 10 goats). A total of 246 blood smears were examined for the presence of intra-erythrocytic inclusions using Giemsa stain. Among the collected specimens, 60 were found positive with an overall prevalence of 24.3%. Among the 60 positive animals, 26 (43.3%) were sheep and 34 (56.7%) were goat. In the peripheral blood samples, the other piroplasmids — Babesia ovis, Theileria ovis, Th. recondita — were followed in an associative form. The ticks were collected, and the species composition was appointed in order to determine the ticks parasitized and have a pathogenic lifestyle in small ruminants. The intensive infection with the ticks of 2 genera — Rhipicephalus and Hyalomma was followed in small ruminants. It was determined that 45.8% of sheep and 35.1% of goats were infected intensively with the ticks of the Hyalomma genus. 110 samples prepared from the ticks of the Hyalomma genus were tested from the PCR test according to the Anaplasma ovis pathogen. 45 samples (40.9%) were assessed positively that 21 samples of them belonged to sheep and 24 of them to goats. 80 samples prepared from the internal organs of the ticks were examined according to the A. ovis parasite and the obtained results were analyzed. In 5 out of 35 samples which detected the parasites, A. ovis was followed, and in 30 samples, the associative parasites: Th. ovis, B. ovis piroplasmids. The Rickettsia and Coxiella pathogens were also detected in the samples.

Molecular Identification of Piroplasmids in Ticks from Infested Small Ruminants in Konya Province, Turkey.

Ticks play a pivotal role in propagating a diverse spectrum of infectious agents that detrimentally affect the health of both humans and animals. In the present study, a molecular survey was executed of piroplasmids in ticks collected from small ruminants in four districts within Konya province, Turkey. Microscopic examination identified 1281 adult ticks, which were categorized into 357 pools based on their species, sexes, host animals, and collection site before DNA extraction. The infection rates were calculated by using a maximum likelihood estimate (MLE) with 95% confidence intervals (CI). Hyalomma detritum, H. excavatum, Rhipicephalus bursa, R. sanguineus, and R. turanicus were identified in this study. Among the five tick species identified here, R. turanicus exhibited the highest infestation rate in both goats and sheep. The presence of Babesia ovis and Theileria ovis based on 18S rRNA was confirmed using molecular assay. The overall MLE of infection rates for B. ovis and T. ovis was 2.49% (CI 1.72-3.46) and 1.46% (CI 0.87-2.23), respectively. The MLE of B. ovis and T. ovis infection rates in R. bursa was 10.80% (CI 7.43-14.90) and 0.33% (CI 0.02-1.42), respectively, while that in R. turanicus was 0.12% (CI 0.01-0.51) and 2.08% (CI 1.25-3.22). This study further confirms that R. turanicus and R. sanguineus can act as vectors for B. ovis, thus advancing our comprehension of tick-borne piroplasmids epidemiology and providing valuable insights for the development of effective control strategies for ticks and tick-borne diseases in Turkey.

Molecular prevalence and associated infection risk factors of tick-borne protozoan and rickettsial blood pathogens in small ruminants

BackgroundTick-borne blood pathogens cause highly pathogenic diseases, which are associated with substantial economic losses in ruminants. Despite this, epidemiological research on these pathogens remains neglected in many countries. This study initiated a regional epidemiological survey that included the detection of molecular prevalence, associated risk factors, and gene sequencing, combined with phylogenetic analysis, targeting the two main tick-borne blood protozoan and rickettsial pathogens of Babesia, Theileria, and Anaplasma that infect small ruminants. One hundred blood samples were collected from 76 sheep and 24 goats.ResultsMicroscopic examination of Giemsa-stained blood films revealed that 73% of the samples were infected with at least one species of the three blood pathogenic organisms. Molecular diagnosis based on the 18 S rRNA for Babesia and Theileria species and the major surface protein 4 (msp4) for Anaplasma species, revealed that 43% of the small ruminants were infected with at least one of these pathogens. The animal’s sex was the most significant associated risk factor, with 49.4% of female animals infected compared with only 4% of male animals (P < 0.05). The open breeding system recorded the highest infection rate for tick-borne blood pathogens. Homology-based and phylogenetic analyses indicated that the specific isolate species were Babesia ovis (B. ovis), Theileria ovis (T. ovis), and Anaplasma ovis (A. ovis), with sequences showing significant identities with isolates from sheep, goats, and other animal species, and geographically diverse countries in Africa, Asia, and Europe, in addition to Egypt.ConclusionThis was the first molecular evidence of B. ovis, T. ovis, and A. ovis infections in sheep and goat populations in the North Coast region of Egypt. More extensive studies are required to develop an epidemiological map of blood pathogenic organisms, while more effective control strategies are required to reduce the burden of tick-borne pathogens on small ruminants.

Evaluation of Oxidative Status, Cytokines, Acute Phase Proteins and Cardiac Damage Markers in Sheep Naturally Infected with Babesia ovis.

Babesiosis is a tick-borne disease caused by protozoon species in the Babesia genus of the Babesiadae family. The systemic inflammatory response induced by infection is considered to be an important feature of the pathophysiology of ovine babesiosis. In this study, it was aimed to determine serum oxidative status, levels of some cytokines, acute phase proteins and heart damage markers in babesiosis infection. A sample of 40 sheep was used for this purpose, of which 20 were healthy and 20 were infected with Babesia ovis. Babesia infection was diagnosed from Giemsa-stained peripheral blood smears. Infection was also confirmed by the polymerase chain reaction (PCR). Sera from blood samples was tested for oxidative stress parameters (malondialdehyde [MDA], total antioxidant status [TAS], superoxide dismutase [SOD], catalase [CAT] and glutathione peroxidase [GPx]), cytokines (interleukins IL-6, IL-1β, IL-10, tumour necrosis factor α (TNF-α) and interferon-ϒ [IFN-ϒ]), acute-phase proteins (C-reactive protein [CRP], serum amyloid A [SAA] and haptoglobin [Hp]) and specific (troponin I [cTnI], creatine kinase-MB [CK-MB]) and nonspecific (lactate dehydrogenase [LDH], aspartate transaminase [AST]) cardiac damage markers. MDA, SOD, CAT, Hp, TAS, IL-6, IL-10, TNF-α, IL-1β, INF-γ, AST, LDH, CK-MB mass and troponin I values were higher in the patient group than in the healthy group (P < 0.05). However, there was not found to be a statistical difference between the healthy and patient groups in terms of GPx, SAA and CRP values (P > 0.05). It can be stated that serum levels of oxidative stress, some acute phase proteins and cardiac damage markers may increase in naturally infected sheep with babesiosis.

Babesia ovis secreted antigen-1 is a diagnostic marker during the active Babesia ovis infections in sheep.

Ovine babesiosis caused by Babesia ovis is an economically significant disease. Recently, a few B. ovis-specific proteins, including recombinant B. ovis secreted antigen-1 (rBoSA1), have been identified. Immunological analyses revealed that rBoSA1 resides within the cytoplasm of infected erythrocytes and exhibits robust antigenic properties for detecting anti-B. ovis antibodies. This protein is released into the bloodstream during the parasite's development. It would be possible to diagnose active infections by detecting this secretory protein. For this purpose, a rBoSA1-specific polyclonal antibody-based sandwich ELISA was optimized in this study. Blood samples taken from the naturally (n: 100) and experimentally (n: 15) infected sheep were analyzed for the presence of native BoSA1. The results showed that native BoSA1 was detectable in 98% of naturally infected animals. There was a positive correlation between parasitemia level in microscopy and protein density in sandwich ELISA. Experimentally infected animals showed positive reactions from the first or second day of inoculations. However, experimental infections carried out by Rhipicephalus bursa ticks revealed the native BoSA1 was detectable from the 7th day of tick attachment when the parasite began to be seen microscopically. Sandwich ELISA was sensitive enough to detect rBoSA1 protein at a 1.52 ng/ml concentration. Additionally, no serological cross-reactivity was observed between animals infected with various piroplasm species, including Babesia bovis, B. bigemina, B. caballi, B. canis, B. gibsoni, Theileria equi, and T. annulata. Taken collectively, the findings show that the rBoSA1-specific polyclonal antibody-based sandwich ELISA can be successfully used to diagnose clinical B. ovis infections in sheep at the early stage.