Skip to main content
Download PDF

Molecular detection and characterization of Anaplasma ovis, Theileria ovis, and Theileria lestoquardi in sheep and goats in Luxor, Egypt

BMC Veterinary Research volume 20, Article number: 260 (2024) Cite this article

Abstract

Background

Tick-borne diseases cause economically significant losses to animal production globally, and anaplasmosis and theileriosis are associated with the greatest losses. However, the spread of the relevant pathogens in flocks of domesticated animals in southern Egypt is little understood. Accordingly, in this study, we aimed to determine the prevalences of Anaplasma ovis, Theileria ovis, and Theileria lestoquardi in southern Egyptian sheep and goats through blood tests, and to make a molecular characterization of the A. ovis detected in sheep targeting a specific gene.

Results

We collected blood samples collected from 300 sheep and goats (n=150 /species) in Luxor Province in southern Egypt, and analyzed them for the presence of A. ovis, T. ovis and T. lestoquardi with screening by conventional and nested PCR targeting the msp4 and msp5, 18S rRNA, and merozoite surface protein genes. For A. ovis 140/300 samples (46.66%) were positive overall, with 90/150 (60%) and 50/150 (33.33%) positive samples in sheep and goats, respectively. Two major surface protein genes of A. ovis, msp4 and msp5, were sequenced using DNA extracted from sheep and goat blood samples, for phylogenetic analysis and genotyping. The msp4 gene sequence revealed no significant genetic diversity, to contrast to data on A. ovis strains from other countries. For T. lestoquardi, 8/150 (5.33%) samples were positive in sheep, but no samples were positive in goats (0%). For T. ovis, 32/150 (21.33%) samples were positive in sheep, but no samples were positive in goats (0%). Sequencing targeting the merozoite surface protein gene for T. lestoquardi and the small subunit ribosomal RNA gene for T. ovis revealed no significant genetic diversity in the study, another contrast to data on A. ovis strains from other countries.

Conclusion

This study provides valuable data on phylogenetic and molecular classifications of A. ovis, T. ovis and T. lestoquardi found in southern Egyptian sheep and goats. It also represents the first report on detection and molecular characterization of T. lestoquardi in southern Egyptian sheep based on the specific merozoite surface protein gene, thus providing valuable data for molecular characterization of this pathogen in southern Egypt.

Peer Review reports

Introduction

Human and livestock health can be threatened by infectious diseases spread by vectors and wildlife [1]. Vector-borne diseases also cause significant economic losses due to high mortality rates and decreased productivity in livestock [2], and vectors spread around 25% of the pathogens that cause concerning diseases in vertebrates [3]. Numerous factors, including globalization and rising international trade, urbanization, climate change, travel, and animal migration, impact the epidemiology and spread of vector-borne diseases [4, 5].

Tick-borne diseases have global economic significance due to the costs associated with animal productivity losses, with anaplasmosis, babesiosis, and tropical theileriosis as diseases that cause some of the greatest economic losses [6]. Anaplasma organisms cause anaplasmosis and are Gram-negative obligate intracellular bacteria that infect multiple hosts. Anaplasmosis is a mild to severe intraerythrocytic disease that causes significant economic losses and changes in the condition of an infected animal rendering it more susceptible to potentially fatal bacterial, viral, or parasitic infections [7].

The most frequent cause of small ruminant anaplasmosis is A. ovis, which is found worldwide. The clinical symptoms of this condition include anorexia, decreased milk production, fever, tiredness, and abortion [8]. Anaplasmosis infection can occur together with other conditions, including stress, poor nutrition, tick infestation, hot weather, and co-infection with various bacteria or parasites. A. ovis infection can be more severe in goats than in sheep, especially in stressed or weak animals, and it is frequently asymptomatic [9]. A. ovis infection is reportedly relatively common and it is not necessarily linked with changes in any health-related variables [10].

Ruminants are important Anaplasma reservoir hosts, and this pathogen can cause persistent infection in animal hosts and cause the spread of the disease [11]. Traditional techniques for ascertaining A. ovis infection in animals include microscopic examination of Giemsa-stained blood smears; this process is not expensive, but it is time consuming, has low sensitivity, and its sensitivity depends on the examiner's skill level [12, 13]. In laboratory and field studies, A. ovis infection has been directly detected using alternative techniques such as PCR and loop-mediated isothermal amplification [14,15,16]. However, the sensitivity of these approaches is constrained, particularly in persistently infected animals, due to the low bacterial levels [17]. The use of serological assays to identify antibodies to A. ovis at all stages of infection has more advantages than other techniques [18].

Theileriosis is a serious hemoprotozoan disease that affects sheep and goats and is caused by pathogenic Theileria species, such as Theileria luwenshuni, Theileria uilenbergi, and T. lestoquardi. Non-pathogenic Theileria spp., such as T.ovis, Theileria separata, and Theileria recondite, frequently infect small ruminants [19,20,21,22]. Malignant theileriosis is caused by T. lestoquardi, which is found in tropical and subtropical areas of Asia, Africa, the Middle East, East and South Europe, and Africa [23, 24]. Following treatment, recovery from acute theileriosis infection is often accompanied by persistence of certain parasites, which is known as the carrier state. In this state, infection is frequently microscopically undetectable, but pathogen transmission may still occur through tick vectors [25, 26]. The presence of the vector in other theileriosis-free locations increases the probability of disease transmission significantly when an infected animal is in the carrier state. Significant reductions in cattle production have been associated with chronic asymptomatic Theileria spp carriage [27]. T. ovis is generally observed in sheep and goats, but it has also been found in water deer [28], dogs, dromedary camels [29, 30], and dairy cows [31]. T. ovis is non-pathogenic to goats and sheep [23], and there have been no clinical cases, so it may easily be overlooked.

Livestock, such as cattle, buffalo, and sheep, are the primary source of meat and milk for most of the population in Egypt, and a major source of income for farmers, accounting for 40% of the agricultural gross domestic product [32]. Anaplasmosis and theileriosis infecting sheep and goats in southern Egypt still have not been definitively profiled genetically, but such genetic profiling is essential for developing control measures against these diseases.

Materials and methods

Collection of samples and clinical examination

The current study focuses on anaplasmosis and theileriosis We targeted local breeds of sheep and goats of various ages and both sexes in the southern part of Egypt, in Luxor Province, for evaluation in the study with blood sample collection at the time of veterinary examinations between January 2021 to January 2022 (Fig.1). Each animal underwent a clinical evaluation before blood sample collection, and its age, sex, body mass index, body temperature, heart rate, respiratory rate, and visible mucous membrane condition were recorded [33].

Fig. 1
figure 1

Map of Egypt indicating the study areas where samples were collected from different regions in Luxor

Full size image

A total of 300 animals were used in this study (150 sheep and 150 goats), of which 120 were male and 180 were female. The animals ranged in age from one to two years, and were classified into age groups (1-year-olds and 2-year-olds, with n=100 and n-200, respectively). Whole blood samples were taken from jugular vein of each animal using clean, sterile vacutainer tubes containing heparin for DNA extraction as a target for PCR amplification. DNA samples were kept at -20°C until analysis.

Detection of pathogens by PCR

All primers used in this study are listed in Table 1 [34,35,36,37], and the PCR conditions are shown in Table 2. Negative controls containing nuclease-free water were used as negative samples. Electrophoresis of PCR products was performed with a 1.5% agarose gel in Tris–acetate–EDTA (TAE) buffer with a Mupid electrophoresis device (Mupid Co., Ltd., Tokyo, Japan), and bands were visualized through a gel documentation system UV device, WUV-M20 (ATTO Co., Ltd., Tokyo, Japan), after being stained with 5 μg /mL ethidium bromide in TAE buffer.

Table 1 The primers for detection of Anaplasma ovis, Theileria ovis and Theileria lestoquardi in sheep and goat
Full size table
Table 2 PCR conditions for the amplification of target fragments of genes of Anaplasma ovis, Theileria ovis and Theileria lestoquardi in sheep and goat
Full size table

DNA extraction and PCR amplification

The 300 blood samples collected in this study (from 150 sheep and 150 goats), were subjected to DNA extraction (from whole blood) using commercial extraction kits (Wizard Genomic DNA Purification Kit, Promega, Madison, WI, USA). A. ovis, T. ovis, and T. lestoquardi were screened in a nested PCR targeting the 18S rRNA T. ovis gene and conventional PCR targeting the msp4 and msp5, and merozoite surface protein genes. The PCR reaction was performed with a total volume of 10 μL using Tks Gflex DNA Polymerase (TaKaRa, Shiga, Japan).

Sequencing and data analysis

A subset of 50 samples was selected from the 300 study samples, and submitted to PCR assays for A. ovis, T. ovis and T. lestoquardi identification targeting the msp4 and msp5, 18S rRNA and merozoite surface protein genes (50 μL aliquots were used for sequence analysis). The amplicons were purified using a NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel, Leicestershire, Duren, Germany) following the manufacturer's protocol. Sequence readings were compared with sequences of reported isolates using BLAST in GenBank. A maximum likelihood phylogenetic tree was constructed using MEGA X software [38], with bootstrap values estimated using 1,000 replicates based on Kimura's two-parameter substitution model [39].

Statistical analysis

The infection rates of A. ovis, T. ovis and T. lestoquardi in sheep and goats were determined by direct counting. Data were evaluated for significant differences and risk factors with a fisher exact probability test. The P value of < 0.05 was considered statistically significant. The 95% confidence intervals were calculated using Vassar Stats software (www.vassarstats.net.).

Results

We detected A. ovis in 140/300 (46.66%) of samples overall, in 90/150 (60%) samples from sheep, and in 50/150 (33.33%) samples from goats (Table 3). A. ovis infection in sheep and goats was evaluated for statically significant associations with a number of risk factors (p < 0.05). By location, the highest prevalence of A. ovis infection was found in northern Luxor province (64 %), followed by the central region (46%) and southern Luxor provinces (40%). The location did not predict infection status when the northern and central regions were compared (p > 0.05); however, it showed significant prediction ability when the southern and northern regions were compared (p < 0.05). A higher infection rate (51%) was found in females than in males (40%), with a significant difference suggesting that sex was a risk factor (p < 0.05). The infection rate was higher in young animals (55%) than in older animals (42.5%). Assessing age as a risk factor, an age not exceeding one year showed a significant difference versus an age not exceeding two years (p < 0.05). Husbandry regime appears to play a role in the greater levels of A. ovis infection, with a lower rate for individual husbandry (37.5%) versus intensive husbandry (65%), and the significant difference suggested that husbandry was a risk factor (p < 0.05) (Table 4)

Table 3 Detection of Anaplasma ovis, Theileria ovis and Theileria lestoquardi infection rate in sheep and goat, from Luxor based on PCR detection in blood samples
Full size table
Table 4 Anaplasma ovis infection rate in sheep and goats in different locations in Luxor
Full size table

We detected T. lestoquardi in 8/150 (5.33%) of samples from sheep, but in no samples from goats (0/150; 0%).). Furthermore, we detected T. ovis in 32/150 (21.33%) of samples from sheep, but never detected in goats (Table 3). The goats in this study were negative for both T. ovis and T. lestoquardi, even though animals from both species in this study were gathered at the same location under both intensive and individual husbandry regimens.

In all A. ovis-positive samples, we detected major surface protein msp4 and msp5 genes, and they were sequenced for phylogenetic analysis and genotyping of this pathogen in the study samples from sheep and goats in Luxor Province in southern Egypt. All sequences were also submitted to GenBank, and the accession numbers shown below can be used to access them.

msp4 gene: OP244840.1, OP244841.1, OP244842.1, and OP244843.1 in goats, and OP244844.1, OP244845.1, OP244846.1, and OP244847.1 in sheep

msp5 gene: OP244852.1, OP244853.1, OP244854.1, and OP244855.1 in goats, and OP244848.1, OP244849.1, OP244850.1, and OP244851.1 in sheep

The phylogenetic analysis for the msp4 gene in sheep and goats in this study compared amplicons from other reported isolates, and alignment results demonstrated 100% identity to sheep from Iran (MH790273.1), Turkey (KY283958.1), and Russia (MT062870.1), and cattle from Russia (MW535731.1); 99.88% identity with sheep from Sudan (KU497698.1) and cattle from China (MN198191.1); 99.75% identity with goats from Mongolia LC412089.1 and LC412084.1; and 99.63% identity with goats from Sudan (KU497709.1), as well as with Anaplasma in Ixodes ricinus from Serbia GQ925819.1. The sequences for the A. ovis msp4 gene were grouped with various isolates from other countries (Fig.2). The phylogenetic analysis for the msp5 gene in sheep and goats in this study compared amplicons separate from other reported isolates with alignment by 99.81% identity to sheep from China GQ483471.1 and by 99.62% identity to sheep from China HM195102.1 (Fig.3).

Fig. 2
figure 2

Phylogenetic relationships of Anaplasma ovis using the maximum likelihood method and Kimura’s 2-parameter model based on major surface protein 4 (msp4) sequences. The percentage of trees in which the associated taxa clustered together is shown next to the branches. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Anaplasma ovis obtained in the present study are represented by green and blue circles

Full size image
Fig. 3
figure 3

Phylogenetic relationships of Anaplasma ovis based on the major surface protein 5 (msp5) sequences, the maximum likelihood method, and Kimura’s 2-parameter model. Next to the branches is a percentage of the trees in which the associated taxa clustered together. Branch lengths are expressed as the number of substitutions per site, and the tree is displayed to scale. Green and blue circles indicate the Anaplasma ovis results from the current study

Full size image

The T. lestoquardi merozoite surface protein sequences established for phylogenetic analysis and genotyping were classified into two groups. One group comprised OP499850.1, OP499851.1 and OP499852.1 and showed 100% identity to the LC430943.1 goat sequence from Iran deposited in GenBank sequences, and maximum identity (99.71%) with the LC430944.1 in sheep from Sudan, and minimum identity (97.16%) with the ON982799.1 in sheep from Pakistan. The second group comprised OP499853.1, OP499854.1, OP499855.1, and showed 100% identity with the sheep sequences from India, MZ074323.1, and ON408247.1, and with sheep sequences from Sudan, KY965146.1 and AF004775.2, but the minimum identity ( 99.04%) was with the MF765610.1 sequence from sheep in Sudan (Fig.4).

Fig. 4
figure 4

Phylogenetic relationships of Theileria lestoquardi using maximum likelihood method and Kimura 2-parameter model based on merozoite surface protein. The percentage of trees in which the associated taxa clustered together is shown next to the branches. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Theileria lestoquardi obtained in the present study were represented by green circles

Full size image

T. ovis small subunit ribosomal RNA was sequenced for phylogenetic analysis and genotyping, the result was submitted to GenBank with accession number from OP389057.1 to OP295071.1 the identical was by 100% with sequences from Egypt MN625903.1, MN625887.1 and MN625886.1 for donkey, boffola and sheep, and also identical by 100% with sheep from Turkey MN493111.1, sheep from Indian MZ220428.1 and sheep from Spain AY533144.1, in addition to it was identical by 99.88% with other sequences in GenBank with Sheep from Iran GU726904.1, Bos taurus from France EU622911.1 and goat from Indian MZ220430.1 (Fig.5).

Fig. 5
figure 5

Phylogenetic relationships of Theileria ovis using maximum likelihood method and Kimura 2-parameter model based on small subunit ribosomal RNA. The percentage of trees in which the associated taxa clustered together is shown next to the branches. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Theileria ovis obtained in the present study were represented by green circles

Full size image

Discussion

The major surface protein 5 is conserved across all known species of Anaplasma marginale, Anaplasma centrale, and A. ovis [40]. Without additional methods such as sequencing and multi-gene amplification, the diagnostic PCRs utilized for A. ovis and A. marginale identification have been unable to distinguish between these pathogens [41]. ELISA and PCR assays have been used to detect A. ovis in animals; however, these assays do not distinguish between different species, and a diagnosis of A. ovis infection is often made when screening samples from sheep [42, 43]. Assays targeting the species-specific msp4 gene have been employed in a number of studies for epidemiological investigations and enabled researchers to clearly differentiate between A. ovis and A. marginale [44]. In North Africa, A. ovis infection has been studied in small ruminants in Egypt, Sudan, Tunisia, and Algeria [42, 45,46,47]. In the current study, the prevalence of A. ovis infection in sheep was 60%, lower than that reported in sheep in other North African countries, specifically, in Sudan at 60.1% [48], and Tunisa at 70.1% [49], and in Mongolia, at 69% [50]. However, our figure for prevalence in sheep exceeded those for Qinghai, China, at 54.5% [51], central and Western Kenya, at 34.2% [52], Turkey, at 29.7% [53], Iran, at 20.8% [54], Pakistan, at 12.5% [55], West Iran, at 10% [56], South Western China, at 5.7% [57], and North East China, at 2.6% [58].

In this study conducted in Egypt’s Luxor province, the rate of A. ovis infection in goats was 33.33%, which is higher than those detected by ELISA in goats in other Egyptian provinces, specifically in Alexandria (23.3%), Behirah (13.8%), Kafr El-Sheikh (22.6%), and El Gharbia (25%) [59].

In terms of age, we found a significant difference between one-year-old (age≤1) and two-year-old (age≤2 years) animals (p < 0.05). This result is consistent with the reportedly higher prevalence of Anaplasma spp infection in sheep below the age of six months [60], although it contrasts with previous reports that adult animals are more susceptible to infection [49, 54].

A higher infection rate (51%) was found in females than in males (40%). The significant difference indicated that sex was a predictive risk factor (p < 0.05), with females appearing more susceptible to A. ovis infection. This susceptibility may be explained by females experiencing greater hormonal fluctuations due to their reproductive cycle [49, 55].

In this study, we also suggested that location could be a predictive risk factor, with a significant difference between animals in the northern and southern regions of Luxor Province (p < 0.05), however, there was no significant difference between animals in the central and northern regions (p > 0.05).

Individual husbandry regime was associated with a lower infection rate (37.5%) than intensive husbandry (65%). The relevant risk factor showed a significant difference (p < 0.05); thus, husbandry may exert an effect whereby animals living in more intensive conditions are exposed to greater numbers of ticks [61]. The msp4 gene sequence analysis in our study apparently did not reveal high genetic diversity for A. ovis, relative to results reported in other countries. The phylogenetic analysis based on this gene in the present study revealed 100% identity with sheep from Iran (MH790273.1), Turkey (KY283958.1), and Russia (MT062870.1), and cattle from Russia (MW535731.1), 99.88% identity with sheep from Sudan (KU4976981) and cattle from China (MN198191.1), 99.75% identity with Mongolian goats (LC412089.1 and LC412084.1), and 99.63% identity with Sudanese goats (KU497709.1), as well as Ixodes ricinus from Serbia GQ925819.1. Understanding the phylogenetic relationships between isolates of A. ovis is vital for informative analysis of intraspecific diversity that could contribute to better prevention and control of anaplasmosis. Therefore, sequencing of the msp4 gene was used to analyze the diversity of our A. ovis samples.

The phylogenetic analysis of the msp5 gene for sheep in this study did not show 100% identity with distinct amplicons from other reported isolates in GenBank. The highest figures were 99.81% identity with Chinese sheep (GQ483471.1) and 99.62% identity with another report in Chinese sheep (HM195102.1). The differences in our sequence were found in two sheep samples in one position, OP244849 and OP244851, in which nucleotide number 106 changed from G to A. This also led to a change in the amino acid from alanine to threonine. These changes are illustrated in Fig. 3 for both sequences in the phylogenetic tree for the msp5 gene. These changes may be due to geographical or ecological factors, tick management, or control practices [16, 62]. Sheep and goats showed high A. ovis infection rates in this study. This may be the result of the limited diagnosis and treatment of sheep and goats with tick, and pasturing these animals in open areas in large numbers thus promoting the quick spread of infection between them. Regular surveys with good data are required to aid farmers and animal health practitioners, and provide guidance in controlling and prioritizing the health of sheep and goats in rural areas.

Theileria lestoquardi is a highly pathogenic ovine and caprine parasite that is believed to be the only Theileria species associated with economic loss present in small ruminants [63,64,65]. The parasite is spread by Hyalomma anatolicum anatolicum, which is found in South-eastern Europe, Northern Africa, Southern Russia, and the Middle East. However, T. lestoquardi distribution has been limited by the range of its vector.

Theileria lestoquardi was detected in a previous study in El-Wady El-Geded Governorate, Egypt, which targeted the 18S ribosomal RNA gene for identification [66]. We consider that our study may be the first research in Egypt to characterize T. lestoquardi based on its merozoite surface protein gene for phylogenetic analysis, and to find 100% identity with reports from goats in Iran, and sheep in India and Sudan.

The results of this study was revealed that the total infection rate of T. lestoquardi was 8/150 (5.33%) in sheep which is higher than a previously reported figure in Egypt (0.87%) [66] but lower than that reported in Oman (10.6%) [67]. We consider that this is the first report on detection of T. lestoquardi based on the specific merozoite surface protein, and the epidemiology of T. lestoquardi needs further investigation in other parts of Egypt to fully elucidate its prevalence. We found a higher T.ovis infection rate, at 32/150 (21.33%), in sheep than previously reported in Egypt (4.37%) [66] and in Oman (2.7%) [67].

In this study, no goats in southern Egypt (Luxor prefecture) were infected with T. lestoquardi or T.ovis, an interesting finding that indicates the need for future research on piroplasmosis in this area. We speculate that T. lestoquardi causes subacute to acute theileriosis in indigenous sheep in the study area, in contrast to the inherent resistance or tolerance displayed by local sheep in other regions where T. lestoquardi is endemic, and that goats are more resistant to the infection than sheep. These variations may reflect parasite epidemiology, seasonal activity of vector ticks, the severity of tick infestation in hosts, behavioral differences between sheep and goats, and the small ruminant population in the studied location [68].

T. ovis primarily infects sheep and goats but has also been detected in water deer (Han et al. 2009), dogs, dromedary camels [29, 30], and dairy cows [31]. T. ovis is non-pathogenic in goats and sheep [23], with no clinical cases of infection having been observed in these species, and it is easily overlooked. T. ovis sequences obtained in the present investigation showed identity (100%) to T. ovis sequences detected in Egyptian donkeys, buffalos, and sheep, and have been grouped in the same clade as sequences of T. ovis from the various regions of Asia, Africa, and Europe. Moreover, these sequences showed strong similarities with corresponding sequences from sheep in the same clade from the Mediterranean area including Turkey, France, Spain, and Iran.

Conclusions

This study focuses on anaplasmosis and theileriosis in local breeds of sheep and goats of various ages and both sexes, and demonstrated higher infection rates for A. ovis in animals under both intensive and individual husbandry regimens. This study represents the first report on the molecular characterization of A. ovis in goats in Egypt. Further research is required to assess the pathogenic potential of these diseases for sheep and goats and to elucidate the evolutionary changes and sources of variation, as information necessary for improvements in livestock health.

Availability of data and material

The partial sequences of the A. ovis msp4 and msp5 genes have been deposited in GenBank with the accession numbers: OP244840.1, OP244841.1, OP244842.1, OP244843.1, OP244844.1, OP244845.1, OP244846.1, OP244847.1, OP244852.1, OP244853.1, OP244854.1, OP244855.1, OP244848.1, OP244849.1, OP244850.1, and OP244851.1.

The partial sequences of the T. lestoquardi merozoite surface protein have been deposited in GenBank with the accession numbers: OP499850.1, OP499851.1, OP499852.1, OP499853.1, OP499854.1, and OP499855.1. The partial sequences of T. ovis small subunit ribosomal RNA deposited in GenBank with the accession numbers from OP389057.1 to OP295071.1.

References

  1. Kuleš J, Potocnakova L, Bhide K, et al. The Challenges and Advances in Diagnosis of Vector-Borne Diseases: Where Do We Stand? Vector Borne Zoonotic Dis. 2017;17(5):285–96. https://doi.org/10.1089/vbz.2016.2074.

    Article  PubMed  Google Scholar 

  2. Lew-Tabor AE, Rodriguez Valle M. A review of reverse vaccinology approaches for the development of vaccines against ticks and tick borne diseases. Ticks Tick Borne Dis. 2016;7(4):573–85. https://doi.org/10.1016/j.ttbdis.2015.12.012.

    Article  CAS  PubMed  Google Scholar 

  3. Bergquist R, Stensgaard AS, Rinaldi L. Vector-borne diseases in a warmer world: Will they stay or will they go? Geospat Health. 2018;13(1):699. https://doi.org/10.4081/gh.2018.699.

    Article  PubMed  Google Scholar 

  4. Harrus S, Baneth G. Drivers for the emergence and re-emergence of vector-borne protozoal and bacterial diseases. Int J Parasitol. 2005;35(11–12):1309–18. https://doi.org/10.1016/j.ijpara.2005.06.005.

    Article  CAS  PubMed  Google Scholar 

  5. Baneth G, Bourdeau P, Bourdoiseau G, et al. Vector-borne diseases–constant challenge for practicing veterinarians: recommendations from the CVBD World Forum. Parasit Vectors. 2012;5:55. https://doi.org/10.1186/1756-3305-5-55.

    Article  PubMed  PubMed Central  Google Scholar 

  6. de Castro JJ. Sustainable tick and tickborne disease control in livestock improvement in developing countries. Vet Parasitol. 1997;71(2–3):77–97. https://doi.org/10.1016/s0304-4017(97)00033-2.

    Article  PubMed  Google Scholar 

  7. Kocan KM, de la Fuente J, Blouin EF, et al. Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia. Parasitology. 2004;129(Suppl):S285–300. https://doi.org/10.1017/s0031182003004700.

    Article  PubMed  Google Scholar 

  8. Stuen S, Longbottom D. Treatment and control of chlamydial and rickettsial infections in sheep and goats. Vet Clin North Am Food Anim Pract. 2011;27(1):213–33. https://doi.org/10.1016/j.cvfa.2010.10.017.

    Article  PubMed  Google Scholar 

  9. Friedhoff KT. Tick-borne diseases of sheep and goats caused by Babesia, Theileria or Anaplasma spp. Parassitologia. 1997;39(2):99–109.

    CAS  PubMed  Google Scholar 

  10. Cabezas-Cruz A, Gallois M, Fontugne M, et al. Epidemiology and genetic diversity of Anaplasma ovis in goats in Corsica, France. Parasit Vectors. 2019;12(1):3. https://doi.org/10.1186/s13071-018-3269-7.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Brown WC, Barbet AF. Persistent Infections and Immunity in Ruminants to Arthropod-Borne Bacteria in the Family Anaplasmataceae. Annu Rev Anim Biosci. 2016;4:177–97. https://doi.org/10.1146/annurev-animal-022513-114206.

    Article  CAS  PubMed  Google Scholar 

  12. Ndung’u LW, Aguirre C, Rurangirwa FR, et al. Detection of Anaplasma ovis infection in goats by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay. J Clin Microbiol. 1995;33(3):675–9. https://doi.org/10.1128/jcm.33.3.675-679.1995.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Silaghi C, Santos AS, Gomes J, et al. Guidelines for the Direct Detection of Anaplasma spp. in Diagnosis and Epidemiological Studies. Vector Borne Zoonotic Dis. 2017;17(1):12–22. https://doi.org/10.1089/vbz.2016.1960.

    Article  PubMed  Google Scholar 

  14. Reisberg K, Selim AM, Gaede W. Simultaneous detection of Chlamydia spp., Coxiella burnetii, and Neospora caninum in abortion material of ruminants by multiplex real-time polymerase chain reaction. J Vet Diagn Invest. 2013;25(5):614–9. https://doi.org/10.1177/1040638713497483.

    Article  PubMed  Google Scholar 

  15. Selim A, Gaede W. Comparative evaluation of PCR assay for direct detection of Mycobacterium avium subsp. paratuberculosis in Ruminant. Asian J Anim, Vet Adv. 2019;10:761–71. https://doi.org/10.3923/ajava.2015.761.771.

    Article  CAS  Google Scholar 

  16. Selim A, Abdelrahman A, Thiéry R, et al. Molecular typing of Coxiella burnetii from sheep in Egypt. Comp Immunol Microbiol Infect Dis. 2019;67: 101353. https://doi.org/10.1016/j.cimid.2019.101353.

    Article  PubMed  Google Scholar 

  17. Shompole S, Waghela SD, Rurangirwa FR, et al. Cloned DNA probes identify Anaplasma ovis in goats and reveal a high prevalence of infection. J Clin Microbiol. 1989;27(12):2730–5. https://doi.org/10.1128/jcm.27.12.2730-2735.1989.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. McGuire TC, Davis WC, Brassfield AL, et al. Identification of Anaplasma marginale long-term carrier cattle by detection of serum antibody to isolated MSP-3. J Clin Microbiol. 1991;29(4):788–93. https://doi.org/10.1128/jcm.29.4.788-793.1991.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ahmed JS, Luo J, Schnittger L, et al. Phylogenetic position of small-ruminant infecting piroplasms. Ann N Y Acad Sci. 2006;1081:498–504. https://doi.org/10.1196/annals.1373.074.

    Article  CAS  PubMed  Google Scholar 

  20. Nagore D, García-Sanmartín J, García-Pérez AL, et al. Identification, genetic diversity and prevalence of Theileria and Babesia species in a sheep population from Northern Spain. Int J Parasitol. 2004;34(9):1059–67. https://doi.org/10.1016/j.ijpara.2004.05.008.

    Article  CAS  PubMed  Google Scholar 

  21. Niu Q, Luo J, Guan G, et al. Detection and differentiation of ovine Theileria and Babesia by reverse line blotting in China. Parasitol Res. 2009;104(6):1417–23. https://doi.org/10.1007/s00436-009-1344-x.

    Article  PubMed  Google Scholar 

  22. Zaeemi M, Haddadzadeh H, Khazraiinia P, et al. Identification of different Theileria species (Theileria lestoquardi, Theileria ovis, and Theileria annulata) in naturally infected sheep using nested PCR-RFLP. Parasitol Res. 2011;108(4):837–43. https://doi.org/10.1007/s00436-010-2119-0.

    Article  PubMed  Google Scholar 

  23. Yin H, Schnittger L, Luo J, et al. Ovine theileriosis in China: a new look at an old story. Parasitol Res. 2007;101(Suppl 2):S191–5. https://doi.org/10.1007/s00436-007-0689-2.

    Article  PubMed  Google Scholar 

  24. Iqbal F, Khattak R, Ozubek S, et al. Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan. Iran J Parasitol. 2013;8(2):289–95.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Al-Fahdi A, Alqamashoui B, Al-Hamidhi S, et al. Molecular surveillance of Theileria parasites of livestock in Oman. Ticks Tick Borne Dis. 2017;8(5):741–8. https://doi.org/10.1016/j.ttbdis.2017.05.008.

    Article  PubMed  Google Scholar 

  26. Uilenberg G. Immunization against diseases caused by Theileria parva: a review. Trop Med Int Health. 1999;4(9):A12–20. https://doi.org/10.1046/j.1365-3156.1999.00446.x.

    Article  CAS  PubMed  Google Scholar 

  27. Gharbi M, Touay A, Khayeche M, et al. Ranking control options for tropical theileriosis in at-risk dairy cattle in Tunisia, using benefit-cost analysis. Rev Sci Tech. 2011;30(3):763-78–78. https://doi.org/10.20506/rst.30.3.2074.

    Article  PubMed  Google Scholar 

  28. Han JI, Jang HJ, Lee SJ, et al. High prevalence of Theileria sp. in wild Chinese Water Deer (Hydropotes inermis argyropus) in South Korea. Vet Parasitol. 2009;164(2–4):311–4. https://doi.org/10.1016/j.vetpar.2009.05.027.

    Article  PubMed  Google Scholar 

  29. Kamani J, Baneth G, Mumcuoglu KY, et al. Molecular detection and characterization of tick-borne pathogens in dogs and ticks from Nigeria. PLoS Negl Trop Dis. 2013;7(3):e2108. https://doi.org/10.1371/journal.pntd.0002108.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Lorusso V, Wijnveld M, Latrofa MS, et al. Canine and ovine tick-borne pathogens in camels Nigeria. Vet Parasitol. 2016;228:90–2. https://doi.org/10.1016/j.vetpar.2016.08.014.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Qi M, Cui Y, Song X, et al. Common occurrence of Theileria annulata and the first report of T. ovis in dairy cattle from Southern Xinjiang, China. Ticks Tick Borne Dis. 2018;9(6):1446–50. https://doi.org/10.1016/j.ttbdis.2018.06.017.

    Article  PubMed  Google Scholar 

  32. Elsify A, Sivakumar T, Nayel M, et al. An epidemiological survey of bovine Babesia and Theileria parasites in cattle, buffaloes, and sheep in Egypt. Parasitol Int. 2015;64(1):79–85. https://doi.org/10.1016/j.parint.2014.10.002.

    Article  PubMed  Google Scholar 

  33. Sherman DM, Robinson RA. Clinical examination of sheep and goats. Vet Clin North Am Large Anim Pract. 1983;5(3):409–26. https://doi.org/10.1016/s0196-9846(17)30052-6.

    Article  CAS  PubMed  Google Scholar 

  34. Ybañez AP, Sivakumar T, Battsetseg B, et al. Specific molecular detection and characterization of Anaplasma marginale in Mongolian cattle. J Vet Med Sci. 2013;75(4):399–406. https://doi.org/10.1292/jvms.12-0361.

    Article  CAS  PubMed  Google Scholar 

  35. de la Fuente J, Atkinson MW, Naranjo V, et al. Sequence analysis of the msp4 gene of Anaplasma ovis strains. Vet Microbiol. 2007;119(2–4):375–81. https://doi.org/10.1016/j.vetmic.2006.09.011.

    Article  CAS  PubMed  Google Scholar 

  36. Altay K, Dumanli N, Holman PJ, et al. Detection of Theileria ovis in naturally infected sheep by nested PCR. Vet Parasitol. 2005;127(2):99–104. https://doi.org/10.1016/j.vetpar.2004.09.012.

    Article  CAS  PubMed  Google Scholar 

  37. Kirvar E, Ilhan T, Katzer F, et al. Detection of Theileria lestoquardi (hirci) in ticks, sheep, and goats using the polymerase chain reaction. Ann N Y Acad Sci. 1998;849:52–62. https://doi.org/10.1111/j.1749-6632.1998.tb11033.x.

    Article  CAS  PubMed  Google Scholar 

  38. Kumar S, Stecher G, Li M, et al. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol Biol Evol. 2018;35(6):1547–9. https://doi.org/10.1093/molbev/msy096.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980;16(2):111–20. https://doi.org/10.1007/bf01731581.

    Article  CAS  PubMed  Google Scholar 

  40. Visser ES, McGuire TC, Palmer GH, et al. The Anaplasma marginale msp5 gene encodes a 19-kilodalton protein conserved in all recognized Anaplasma species. Infect Immun. 1992;60(12):5139–44. https://doi.org/10.1128/iai.60.12.5139-5144.1992.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Torina A, Agnone A, Blanda V, et al. Development and validation of two PCR tests for the detection of and differentiation between Anaplasma ovis and Anaplasma marginale. Ticks Tick Borne Dis. 2012;3(5–6):283–7. https://doi.org/10.1016/j.ttbdis.2012.10.033.

    Article  PubMed  Google Scholar 

  42. Renneker S, Abdo J, Salih DE, et al. Can Anaplasma ovis in small ruminants be neglected any longer? Transbound Emerg Dis. 2013;60(Suppl 2):105–12. https://doi.org/10.1111/tbed.12149.

    Article  PubMed  Google Scholar 

  43. Mason KL, Gonzalez MV, Chung C, et al. Validation of an improved Anaplasma antibody competitive ELISA for detection of Anaplasma ovis antibody in domestic sheep. J Vet Diagn Invest. 2017;29(5):763–6. https://doi.org/10.1177/1040638717709494.

    Article  CAS  PubMed  Google Scholar 

  44. Liu Z, Peasley AM, Yang J, et al. The Anaplasma ovis genome reveals a high proportion of pseudogenes. BMC Genomics. 2019;20(1):69. https://doi.org/10.1186/s12864-018-5374-6.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Belkahia H, Ben Said M, El Mabrouk N, et al. Seasonal dynamics, spatial distribution and genetic analysis of Anaplasma species infecting small ruminants from Northern Tunisia. Infect Genet Evol. 2017;54:66–73. https://doi.org/10.1016/j.meegid.2017.06.016.

    Article  CAS  PubMed  Google Scholar 

  46. Belkahia H, Ben Said M, El Hamdi S, et al. First molecular identification and genetic characterization of Anaplasma ovis in sheep from Tunisia. Small Rumin Res. 2017;121:404–10. https://doi.org/10.1016/j.smallrumres.2014.07.009.

    Article  Google Scholar 

  47. Aouadi A, Leulmi H, Boucheikhchoukh M, et al. Molecular evidence of tick-borne hemoprotozoan-parasites (Theileria ovis and Babesia ovis) and bacteria in ticks and blood from small ruminants in Northern Algeria. Comp Immunol Microbiol Infect Dis. 2017;50:34–9. https://doi.org/10.1016/j.cimid.2016.11.008.

    Article  PubMed  Google Scholar 

  48. Lee SH, Mossaad E, Ibrahim AM, et al. Detection and molecular characterization of tick-borne pathogens infecting sheep and goats in Blue Nile and West Kordofan states in Sudan. Ticks Tick Borne Dis. 2018;9(3):598–604. https://doi.org/10.1016/j.ttbdis.2018.01.014.

    Article  PubMed  Google Scholar 

  49. Belkahia H, Said MB, El Hamdi S, Yahiaoui M, Gharbi M, Daaloul-Jedidi M, et al. First molecular identification and genetic characterization of Anaplasma ovis in sheep from Tunisia Small. Rumin Res. 2014;121:404–10. https://doi.org/10.1016/j.smallrumres.2014.07.009.

    Article  Google Scholar 

  50. Enkhtaivan B, Narantsatsrala S, Davaasurena B, Otgonsurena D, Amgalanbaatara T, Uuganbayara E, et al. Molecular detection of Anaplasma ovis in small ruminants and ixodid ticks from Mongolia. Parasitol Int. 2019;69:47–53. https://doi.org/10.1016/j.parint.2018.11.004.

    Article  CAS  PubMed  Google Scholar 

  51. Zhang X, Liu Z, Yang J, Chen Z, Guan G, Ren Q, et al. Multiplex PCR for diagnosis of Theileria uilenbergi, Theileria luwenshuni, and Theileria ovis in small ruminants. Parasitol Res. 2014;113:527–31. https://doi.org/10.1007/s00436-013-3684-9.

    Article  PubMed  Google Scholar 

  52. Ringo AE, Aboge GO, Adjou Moumouni PF, Hun Lee S, Jirapattharasate C, Liu M, et al. Molecular detection and genetic characterisation of pathogenic Theileria, Anaplasma and Ehrlichia species among apparently healthy sheep in central and western Kenya. Onder J Vet Res. 2019;86:1630. https://doi.org/10.4102/ojvr.v86i1.1630.

    Article  CAS  Google Scholar 

  53. Zhou M, Cao S, Sevinc F, Sevinc M, Ceylan O, Ekici S, et al. Molecular detection and genetic characterization of Babesia, Theileria and Anaplasma amongst apparently healthy sheep and goats in the central region of Turkey. Tick Tick Born Dis. 2017;8:246–52. https://doi.org/10.1016/j.ttbdis.2016.11.006.

    Article  Google Scholar 

  54. Yousefi A, Rahbari S, Shayan P, Sadeghi-dehkordi Z, Bahonar A. Molecular detection of Anaplasma marginale and Anaplasma ovis in sheep and goat in west highland pasture of Iran. Asia Pac J Trop Biomed. 2017;7:455–9. https://doi.org/10.1016/j.apjtb.2017.01.017.

    Article  Google Scholar 

  55. Naeem M, Amaro-Estrada I, Taqadus A, Swelum AA, Alqhtani AH, Asif M, Sajid M, Khan AU, Tariq A, Anjum S, Khan A, Iqbal F. Molecular prevalence and associated risk factors of Anaplasma ovis in Pakistani sheep. Front Vet Sci. 2023;29(10):1096418. https://doi.org/10.3389/fvets.2023.1096418.PMID:37065244;PMCID:PMC10095557.

    Article  Google Scholar 

  56. Mohammadian B, Noaman V, Emami SJ. Molecular survey on prevalence and risk factors of Anaplasma spp. infection in cattle and sheep in West of Iran. Trop Anim Health Prod. 2021;53:266. https://doi.org/10.1007/s11250-021-02707-1.

    Article  PubMed  Google Scholar 

  57. Yang J, Hana R, Niua Q, Liua Z, Guana G, Liua G, et al. Occurrence of four Anaplasma species with veterinary and public health significance in sheep Northwestern China. Tick Tick Born Dis. 2017;9:82–5. https://doi.org/10.1016/j.ttbdis.2017.10.005.

    Article  Google Scholar 

  58. Shi Y, Yang J, Guan G, Liu Z, Luo J, Song M. Molecular investigation of Anaplasma species in sheep from Heilongjiang Province, northeast China identified four Anaplasma species and a novel genotype of Anaplasma capra. Parasitol Int. 2020;76:102072. https://doi.org/10.1016/j.parint.2020.102072.

    Article  CAS  PubMed  Google Scholar 

  59. Selim A, Attia KA, Alsubki RA, et al. The first study on the seroprevalence of Anaplasma spp. in small ruminants and assessment of associated risk factors in North Egypt. Vet World. 2022;15(5):1221–7. https://doi.org/10.14202/vetworld.2022.1221-1227.

    Article  PubMed  PubMed Central  Google Scholar 

  60. Niaz S, Ur Rahman Z, Ali I, et al. Molecular prevalence, characterization and associated risk factors of Anaplasma spp. and Theileria spp. in small ruminants in Northern Pakistan. Parasite. 2021;28:3. https://doi.org/10.1051/parasite/2020075.

    Article  PubMed  PubMed Central  Google Scholar 

  61. Cutler SJ, Fooks AR, van der Poel WH. Public health threat of new, reemerging, and neglected zoonoses in the industrialized world. Emerg Infect Dis. 2010;16(1):1–7. https://doi.org/10.3201/eid1601.081467.

    Article  PubMed  PubMed Central  Google Scholar 

  62. Abdullah H, Amanzougaghene N, Dahmana H, et al. Multiple vector-borne pathogens of domestic animals in Egypt. PLoS Negl Trop Dis. 2021;15(9): e0009767. https://doi.org/10.1371/journal.pntd.0009767.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Leemans I, Fossum C, Johannisson A, et al. Comparative studies on surface phenotypes of Theileria lestoquardi and T. annulata schizont-infected cells. Parasitol Res. 2001;87(9):768–77. https://doi.org/10.1007/s004360100388.

    Article  CAS  PubMed  Google Scholar 

  64. Li Y, Chen Z, Liu Z, et al. Molecular identification of Theileria parasites of northwestern Chinese Cervidae. Parasit Vectors. 2014;7:225. https://doi.org/10.1186/1756-3305-7-225.

    Article  PubMed  PubMed Central  Google Scholar 

  65. El-Alfy ES, Abbas I, Baghdadi HB, El-Sayed SAE, Ji S, Rizk MA. Molecular Epidemiology and Species Diversity of Tick-Borne Pathogens of Animals in Egypt: A Systematic Review and Meta-Analysis. Pathogens. 2022;11(8):912. Published 2022 Aug 14. https://doi.org/10.3390/pathogens11080912

  66. Al-Hosary AA, ElSify A, Salama AA, et al. Phylogenetic study of Theileria ovis and Theileria lestoquardi in sheep from Egypt: Molecular evidence and genetic characterization. Vet World. 2021;14(3):634–9. https://doi.org/10.14202/vetworld.2021.634-639

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Al-Hamidhi S, Elshafie EI, Yaghfoori S, et al. A comparative study of single Theileria lestoquardi and mixed infections with Theileria ovis. Parasit Vectors. 2021;14(1):370. https://doi.org/10.1186/s13071-021-04864-6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Inci A, Ica A, Yildirim A, Düzlü Ö. Identification of Babesia and Theileria species in small ruminants in Central Anatolia (Turkey) via reverse line blotting. T J Vet Anim Sci. 2010;34:205–10.

    CAS  Google Scholar 

Download references

Acknowledgement

The authors would like to express their gratitude to Henry Smith, the Joint Faculty of Veterinary Medicine, Kagoshima University, Japan, for his support and assistance in the English edition of this work.

Funding

Open access funding provided by The Science, Technology & Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB). This work was supported by the Arab Republic of Egypt, Ministry of Higher Education and Scientific Research (Faculty of Veterinary Medicine, South Valley University) in the form of a scholarship as a post-doctor researcher for H.Y.A.H.M in the laboratory of infectious diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, Japan.

Author information

Authors and Affiliations

  1. Division of Infectious Diseases, Animal Medicine Department, Faculty of Veterinary Medicine, South Valley University, Qena, 83523, Egypt

    Hassan Y. A. H. Mahmoud & Alsagher O. Ali

  2. Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, 890-0065, Japan

    Tetsuya Tanaka

  3. Department of Fish Diseases and Management, Faculty of Veterinary Medicine, South Valley University, Qena, 83523, Egypt

    Walaa F. A. Emeish

Authors
  1. Hassan Y. A. H. Mahmoud
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tetsuya Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alsagher O. Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Walaa F. A. Emeish
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

H.Y.A.H.M. and T.T., conceptualization and design experiments, formal analysis, investigation: H.Y.A.H.M., and T.T.; resources and shared materials: H.Y.A.H.M., W.F.E., A.O.A., and T.T.; writing the original draft: H.Y.A.H.M. and T.T.; writing—review and editing: H.Y.A.H.M., W.F.E., A.O.A., and T.T.; project administration and funding acquisition: H.Y.A.H.M. and T.T. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Hassan Y. A. H. Mahmoud.

Ethics declarations

Ethics approval and consent to participate

This research was performed after obtaining verbal informed consent from the animal owners and the Ethics approval in this study was approved by the Ethical Research Committee at the Faculty of Veterinary Medicine, South Valley University (No. 10/09.02.2021, and 52/13.9.2022). All methods were carried out in accordance with relevant guidelines and regulations.

Consent to publication

Not applicable.

Competing interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mahmoud, H.Y.A.H., Tanaka, T., Ali, A.O. et al. Molecular detection and characterization of Anaplasma ovis, Theileria ovis, and Theileria lestoquardi in sheep and goats in Luxor, Egypt. BMC Vet Res 20, 260 (2024). https://doi.org/10.1186/s12917-024-04109-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12917-024-04109-5

Keywords

BMC Veterinary Research

ISSN: 1746-6148

Contact us