Egyptian Cattle Research Articles

Integrated Characterization and Pathogen Detection of Ticks in Egyptian Cattle and Equines: Focus on Rhipicephalus annulatus and Hyalomma excavatum

Integrated Characterization and Pathogen Detection of Ticks in Egyptian Cattle and Equines: Focus on Rhipicephalus annulatus and Hyalomma excavatum

Gut Microbiota Diversity of Local Egyptian Cattle Managed in Different Ecosystems.

The animal gastrointestinal tract contains a complex microbiome whose composition ultimately reflects the co-evolution of microorganisms with their animal host and their host's environment. This study aimed to gain insights into the adaptation of the microbiota of local Egyptian cattle to three different ecosystems (Upper Egypt, Middle Egypt, and Lower Egypt) distributed across 11 governorates (with an average of 12 animals per governorate) using amplicon sequencing. We analyzed the microbiota from 136 fecal samples of local Egyptian cattle through a 16S rRNA gene sequencing approach to better understand the fecal microbial diversity of this breed which developed under different ecosystems. An alpha diversity analysis showed that the fecal microbiota of the Egyptian cattle was not significantly diverse across areas, seasons, sexes, or farm types. Meanwhile, microbiota data revealed significant differences in richness among age groups (p = 0.0018). The microbial community differed significantly in the distribution of its relative abundance rather than in richness across different ecosystems. The taxonomic analysis of the reads identified Firmicutes and Actinobacteriota as the dominant phyla, accounting for over 93% of the total bacterial community in Egyptian cattle. Middle Egypt exhibited a different microbial community composition compared to Upper and Lower Egypt, with a significantly higher abundance of Firmicutes and Euryarchaeota and a lower abundance of Actinobacteriota in this region than the other two ecosystems. Additionally, Middle Egypt had a significantly higher relative abundance of the Methanobacteriaceae family and the Methanobrevibacter genera than Lower and Upper Egypt. These results suggest a difference in the adaptation of the fecal microbial communities of Egyptian cattle raised in Middle Egypt. At the genus level, eleven genera were significantly different among the three ecosystems including Bacillus, DNF00809, Kandleria, Lachnospiraceae_NK3A20_group, Methanobrevibacter, Mogibacterium, Olsenella, Paeniclostridium, Romboutsia, Turicibacter, and UCG-005. These significant differences in microbiota composition may impact the animal's adaptation to varied environments.

First characterization of major histocompatibility complex class II DRB3 diversity in cattle breeds raised in Egypt

Genes encoding bovine leukocyte antigen (BoLA) enable the immune system to identify pathogens. Therefore, these genes have been used as genetic markers for infectious and autoimmune diseases as well as for immunological traits in cattle. Although BoLA polymorphisms have been reported in various cattle breeds worldwide, they have not been studied in cattle populations in Egypt. In this study, we characterized BoLA-DRB3 in two local Egyptian populations and one foreign population using polymerase chain reaction-sequence-based typing (PCR-SBT) method. Fifty-four previously reported BoLA-DRB3 alleles and eight new alleles (BoLA-DRB3*005:08, *015:07, *016:03, *017:04, *020:02:02, *021:03, *164:01, and *165:01) were identified. Alignment analysis of the eight new alleles revealed 90.7–98.9 %, and 83.1–97.8 % nucleotide and amino acid identities, respectively, with the BoLA-DRB3 cDNA clone NR-1. Interestingly, BoLA-DRB3 in Egyptian cattle showed a high degree of allelic diversity in native (na = 28, hE > 0.95), mixed (na = 61, hE > 0.96), and Holstein (na = 18, hE > 0.88) populations. BoLA-DRB3*002:01 (14.3 %), BoLA-DRB3*001:01 (8.5 %), and BoLA-DRB3*015:01 (20.2 %) were the most frequent alleles in native, mixed, and Holstein populations, respectively, indicating that the genetic profiles differed in each population. Based on the allele frequencies of BoLA-DRB3, genetic variation among Egyptian, Asian, African, and American breeds was examined using Nei’s distances and principal component analysis. The results suggested that native and mixed cattle populations were most closely associated with African breeds in terms of their gene pool, whereas Holstein cattle were more distinct from the other breeds and were closely related to Holstein cattle populations from other countries.

Effect of water quality on causes of calf mortality in cattle-farm-associated epidemics

Abstract. Poor-quality drinking water plays a detrimental role in the suppression of calf immunity, giving rise to an increased rate of calf mortality. The present study aims to evaluate the causes of calf mortality in beef and dairy farms in relation to drinking water quality (DWQ). A convenience sample of 132 Egyptian cattle farms suffering from emerging epidemics was surveyed by collecting drinking water samples for physicochemical and microbial analysis and using a questionnaire to record hygienic risk factors affecting calf health. Statistical analysis correlates water parameters with rates of calf diarrhea, respiratory problems, severe depression, sudden death and mortality. High percentages of water sample quality parameters, e.g. pH, total dissolved solids (TDSs), hardness, chloride, nitrate, sulfate, total colony count (TCC) and total coliform count (TCFC), are above permissible limits. Water parameters, except pH, show a significant moderate positive correlation with causes of calf mortality (ρ 0.331–0.66) in winter and summer. Each cause of calf mortality was predicted by a specific water parameter, and the water nitrate level was the highest predictor, with the highest values (β = 0.504–0.577), followed by the water TDS, sulfate and microbial levels. Weak to moderate correlation (ρ 0.151–0.367) was found between calf mortality causes and some hygienic risk factors such as operation type, calf housing, calf feeders, bedding type, water source, water pipe type, drinker lining and wheel dipping. We could conclude that DWQ greatly affects causes of calf mortality, but we cannot exclude some farm hygienic risk factors.

Neospora caninum infection in dairy cattle in Egypt: a serosurvey and associated risk factors

Neospora caninum (N. caninum) is one of the causative agents that causing cattle abortion, and severe economic losses. Due to the scarcity of data on N. caninum infection in Egyptian cattle, the purpose of this study was to estimate the seroprevalence and determine the risk factors for parasite infection. In four governorates in northern Egypt, 540 blood samples from cattle were taken, and tested using a commercial ELISA kit. The overall seroprevalence of N. caninum in examined cattle was 28.89%. A multivariate logistic regression model determined that age (OR = 2.63, P < 0.001), manual milking (OR = 1.39, P = 0.14), abortion history (OR = 2.78, P < 0.0001), repetition of estrus (OR = 2.31, P < 0.0001), and contact with dogs (OR = 2.57, P < 0.0001) were significant risk factors. The findings proved that N. caninum infection was one of the factors contributing to abortion and financial losses in dairy cattle in Egypt. Therefore, the application of sanitary security and control programs is very important in dairy farms.

The influence of risk factors on bovine leukemia virus infection and proviral load in egyptian cattle.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leucosis (EBL), which affects cattle globally. In Egypt, BLV control strategies have been ignored because of the shortage of BLV research studies and the silent infection in most animals. This study aimed to identify the risk factors associated with the prevalence of BLV among dairy and beef cattle from six different geographic and climatic provinces in Egypt. Additionally, risk factors affecting the BLV proviral load (PVL) among the positive cattle were targeted. The total BLV prevalence in cattle from six investigated Egyptian provinces was 24.2% (105/433), while the mean PVL (8651.6 copies /105 white blood cells) was absolutely high as estimated by the BLV-CoCoMo-quantitative polymerase chain reaction (qPCR)-2 assay. Analysis of the influence of risk factors (age, sex, breed, production type, farm size, and location) on BLV prevalence indicated that the Holstein breed (OR = 1.582, p = 0.007), beef cattle (OR = 1.088, p = 0.0001), large-size farms (OR = 1.26, p = 0.0001), and cattle from Damietta (OR = 1.43, p = 0.0001) and Cairo (OR = 1.16, p = 0.0001) were ultimately proven the most important risks for BLV infection. The risk factors were analyzed considering the BLV PVL levels in the BLV-positive cases. Significantly high PVL (HPVL) levels were observed in cattle > 5 years old (p < 0.0001), females (p = 0.0008), Holstein (p < 0.0001), dairy cows (p = 0.0053), large-size farms (p < 0.0001), and cattle from Damietta (p < 0.0001) compared to other categories. Contrary, no significant differences in PVL levels were reported between the Native and Mixed cattle breeds (p = 0.13). Ultimately, the logistic regression model indicated that the probability of carrying HPVL in cattle > 5 years is 1.27 (95% CI: 1.03-2.09, p < 0.001) times more likely compared to cattle < 2 years old. In conclusion, the findings were valuably correlating the BLV prevalence with PVL as an indicator of the risk of BLV infection.

Bluetongue virus infection in cattle: serosurvey and its associated risk factors.

Bluetongue virus (BTV) is a vector-borne virus that primarily affects sheep. However, the disease is usually asymptomatic in cattle without obvious clinical signs related to BTV infection. Although there is evidence of BTV antibodies through serology in Egypt, it is still unknown whether Egyptian cattle have ever been exposed to the virus in the north or south of the country. The study's aims were to determine the seroprevalence of BTV and evaluate the potential risk factors for BTV infection in cattle in Egypt. We used a competitive ELISA to screen 690 healthy cattle for BTV-specific immunoglobulin G (IgG) antibodies in four governorates in Egypt. A total seroprevalence of BTV antibodies in examined cattle was 51.47%, 95%CI: 48.01-55.45. The odds of BTV seropositivity were higher in Aswan (OR=1.30, 95%CI: 0.71-2.36), females (OR=3.29, 95%CI: 1.87-5.79), and elder cattle >8 years (OR=12.91, 95%CI: 6.63-25.13). Moreover, cattle contacted with other animals (OR=1.40, 95%CI: 0.94-2.10), with history of abortion (OR=4.88, 95%CI: 3.14-7.59), and those living with presence of insects (OR=12.34, 95%CI: 8-19.30) were more likely to be infected with bluetongue (BT). To effectively predict and respond to a potential BTV outbreak in Egypt, surveillance for BTV infection should be expanded to cover other susceptible ruminants and the range of the insect vectors.

Molecular Prevalence and Risk Factors Analysis of Asymptomatic Infection with Anaplasma marginale and Theileria annulata in Egyptian Cattle

Molecular Prevalence and Risk Factors Analysis of Asymptomatic Infection with Anaplasma marginale and Theileria annulata in Egyptian Cattle

The First Serological Detection and Risk Factors Analysis of Akabane Virus in Egyptian Cattle.

Akabane virus (AKAV) is an insect-borne virus belonging to the genus Orthobunyavirus of the family Peribunyaviridae. It is the etiologic agent of Akabane disease (AD), which emerged in Asia, Australia, and the Middle East causing severe economic losses among domestic and wild animals. AKAV has not received enough attention in Egypt, and its evidence among Egyptian animals has never been reported. Therefore, this study used ELISA assay to investigate the seroprevalence of AKAV among Egyptian dairy and beef cattle in eight localities of Beheira province, north Egypt. Out of 368 investigated plasma samples, the overall AKAV seroprevalence was 54.3% (95% CI: 50.8-61.4). AKAV antibodies were detected in all examined cattle farms (7/7) and the majority of abattoirs (8/9). Age, sex, breed, and location of the tested cattle were analyzed as risk factors for AKAV infection. A higher significant increase in seropositivity was obtained in cattle who were aged >5 years (p < 0.0001; OR = 9.4), females (p < 0.0001, OR = 8.3), or Holstein breed (p < 0.0001, OR = 22.6) than in younger ages, males, and Mixed and Colombian zebu breeds, respectively. Moreover, a significant variation in AKAV seroprevalence between the tested locations was noticed. Ultimately, a multivariable analysis concluded that age (p = 0.002, OR = 3.32, 95% CI = 1.57-7.04) and breed (p = 0.03, OR = 1.69, 95% CI = 1.05-2.72) were significant risks for AKAV infection. In conclusion, this study is the first to detect AKAV infection in Egyptian animals.

The First Molecular Survey on Bovine Immunodeficiency Virus in Egyptian Cattle

The First Molecular Survey on Bovine Immunodeficiency Virus in Egyptian Cattle

Identification and characterisation of lumpy skin disease virus recently isolated from Giza, Egypt

Lumpy skin disease (LSD) is a viral disease, geographically distributed in Africa and now, vigorously spread in the Near East and also in Europe and Asia. It has a significant economic impact on cattle industry in Africa. The aim of this study was isolation and rapid identification of LSD virus circula­ting in Egypt from clinically suspected cattle based on clinical and molecular basis in a rapid and accurate way. Fifteen representative specimens (skin sitfasts) were collected in 2018 from clinically infected cattle in Giza governorate, Egypt. The virus was isolated on chorioallantoic membrane of specific pathogen free embryonated chicken eggs and Madin Darby Bovine Kidney tissue culture cells. The isolated virus was identified and confirmed by conventional polymerase chain reaction (PCR) and real-time PCR. Histopathological examination of the lesions showed a pathognomic intracytoplasmic inclusion body in dermal stroma section. The section of dermal layer revealed vasculitis with projection of its endothelial lining. It was concluded that LSD was enzootic in Egypt and still circulating among Egyptian cattle so that LSD virus could be isolated and identified by traditional and molecular diagnostic methodes. Real time PCR assay could be applied for rapid and accurate confirmation of the field isolate of LSD virus.

Morphological and ultrastructural features of the laryngeal mound of Egyptian Cattle Egret (Bubulcus ibis, Linnaeus, 1758)

BackgroundAccording to our acknowledgment this is the first full anatomical description of the studied laryngeal mound of the Egyptian Cattle Egret (Bubulcus ibis, Linnaeus, 1758). This study was obtained with the aid of scanning electron microscopy (SEM) and histological techniques. Heads of ten healthy adult male Egyptian cattle egrets were used in this study.ResultsThe laryngeal mound (Mons laryngealis) was a pear-shaped musculoskeletal elevation. It represented 20.55 and 67.87% of the total length of the oropharyngeal floor and the pharyngeal floor, respectively. By SEM view, the lateral aspect of the caudal third of the laryngeal mound had a serrated mucosal appearance, forming of 6–7 finger-like projections. Furthermore, the terminal part of the laryngeal mound (except the middle part) was bordered a transverse row of pyramidal-shaped papillae, which demarcated from the esophagus. Histologically, laryngeal salivary glands termed (cricoarytenoid salivary glands) of the laryngeal mound were simple tubular type and were arranged in one row within the lamina propria connective tissue close to the lamina epithelialis. Those glands were surrounded by abundant aggregation of lymphocytes, extended overlying the surface lining epithelium. The glottis within the laryngeal mound was supported by hyaline cartilages; dorsally by paired arytenoid cartilages, ventrolaterally by cricoid cartilage, and caudodorsally by procricoid cartilage. Two groups of intrinsic laryngeal skeletal muscles have connected the cartilages. The glandular epithelium of the laryngeal salivary glands and chondrocytes of the laryngeal cartilages showed strongly positive alcian blue reaction.ConclusionsThe laryngeal mound shows certain features that are unique as an adaptation to lifestyles and bird’s habitat.

Genetic characterization of upper respiratory tract virome from nonvaccinated Egyptian cow-calf operations.

Bovine respiratory disease (BRD) is the costliest complex disease affecting the cattle industry worldwide, with significant economic losses. BRD pathogenesis involves several interactions between microorganisms, such as bacteria and viruses, and management factors. The present study aimed to characterize the nasal virome from 43 pooled nasal swab samples collected from Egyptian nonvaccinated cow-calf operations with acute BRD from January to February 2020 using metagenomic sequencing. Bovine herpesvirus-1 (BHV-1), first detection of bovine herpesvirus-5 (BHV-5), and first detection of bovine parvovirus-3 (BPV-3) were the most commonly identified in Egyptian cattle. Moreover, phylogenetic analysis of glycoprotein B revealed that the BHV-1 isolate is closely related to the Cooper reference strain (genotype 1.1), whereas the BHV-5 isolate is closely related to the reference virus GenBank NP_954920.1. In addition, the whole-genome sequence of BPV-3 showed 93.02% nucleotide identity with the reference virus GenBank AF406967.1. In this study, several DNA viruses, such as BHV-1 and first detection BHV-5, and BPV-3, were detected and may have an association with the BRD in Egyptian cattle. Therefore, further research, including investigating more samples from different locations to determine the prevalence of detected viruses and their contributions to BRD in cattle in Egypt, is needed.

Isolation and Genetic Characterization of Hobi- Like Pestivirus Circulating in Egyptian Cattle during 2019-2020

Isolation and Genetic Characterization of Hobi- Like Pestivirus Circulating in Egyptian Cattle during 2019-2020

Do Pharaohs’ cattle still graze the Nile Valley? Genetic characterization of the Egyptian Baladi cattle breed

Egyptian Baladi cattle is one of the few native taurine breeds which remain in the original domestication area of Bos taurus, the Nile Valley (Upper Egypt). Apart from its evolutive relevance, the breed is well-adapted and integrated into the traditional family farming systems, with great potentialities for local sustainable rural development. The diversity and structure of the Baladi population were assessed using 28 genetic microsatellite markers. Within and between-breeds diversity, its relations with other cattle breeds, and an inference on its origin and evolution, were analyzed. Results reported high levels of diversity, with an average number of alleles (Na) of 11.39, observed heterozygosity (Ho) of 0.68, and expected heterozygosity (He) of 0.75. The studied population was in Hardy-Weinberg disequilibrium, which could be ascribed to its internal structure. The comparison between breeds using factorial correspondence analysis, Reynolds genetic distance evaluation, and genetic structure analysis suggested slight influences of Bos indicus. Upper Egypt’s rural communities have preserved the Baladi cattle breed, protecting this historical and biological heritage against the constant and indiscriminate introduction of exotic genetic resources along the history of development of the breed. Developing and implementing conservation and breeding programs is crucial for the preservation and improvement of the breed. HIGHLIGHTS Baladi cattle breed is a well-defined and highly diverse breed. Higher diversity levels are found in northern and central governorates. A clear geographical cluster is formed around the Nile Delta. Baladi cattle may be reminiscent of the ancient ‘pharaoh’ cattle. Assiut’s population acted as the most ancestral population nucleus.

Characterisation of S. aureus/MRSA CC1153 and review of mobile genetic elements carrying the fusidic acid resistance gene fusC

While many data on molecular epidemiology of MRSA are available for North America, Western Europe and Australia, much less is known on the distribution of MRSA clones elsewhere. Here, we describe a poorly known lineage from the Middle East, CC1153, to which several strains from humans and livestock belong. Isolates were characterised using DNA microarrays and one isolate from the United Arab Emirates was sequenced using Nanopore technology. CC1153 carries agr II and capsule type 5 genes. Enterotoxin genes are rarely present, but PVL is common. Associated spa types include t504, t903 and t13507. PVL-positive CC1153-MSSA were found in Egyptian cattle suffering from mastitis. It was also identified among humans with skin and soft tissue infections in Saudi Arabia, France and Germany. CC1153-MRSA were mainly observed in Arabian Gulf countries. Some isolates presented with a previously unknown SCCmec/SCCfus chimeric element in which a mec B complex was found together with the fusidic acid resistance gene fusC and accompanying genes including ccrA/B-1 recombinase genes. Other isolates carried SCCmec V elements that usually also included fusC. Distribution and emergence of CC1153-MRSA show the necessity of molecular characterization of MRSA that are resistant to fusidic acid. These strains pose a public health threat as they combine resistance to beta-lactams used in hospitals as well as to fusidic acid used in the community. Because of the high prevalence of fusC-positive MRSA in the Middle East, sequences and descriptions of SCC elements harbouring fusC and/or mecA are reviewed. When comparing fusC and its surrounding regions from the CC1153 strain to available published sequences, it became obvious that there are four fusC alleles and five distinct types of fusC gene complexes reminiscent to the mec complexes in SCCmec elements. Likewise, they are associated with different sets of ccrA/B recombinase genes and additional payload that might include entire mec complexes or SCCmec elements.

Lumpy skin disease outbreaks investigation in Egyptian cattle and buffaloes: Serological evidence and molecular characterization of genome termini

Lumpy skin disease (LSD) is an endemic highly infectious viral disease affecting cattle in Egypt. This study aimed to identify and characterize the LSD virus (LSDV) outbreaks in Egypt between 2016 and 2018 and to determine the role of Egyptian buffaloes in the epidemiology of LSD. A total of 44 skin biopsies (41 from cattle and 3 from buffaloes) and 31 blood samples from asymptomatic buffaloes in contact with clinically infected cattle were collected from 7 Egyptian governorates and tested by real-time (rt)-PCR. The positive samples were further isolated, and the isolates were analyzed by conventional PCR to amplify the LSDV001 and LSDV002 genes; three isolates were sequenced, and the phylogenetic tree was constructed. In addition, 198 serum samples (102 from cattle and 96 from contact buffaloes) were examined using ELISA. Out of 44 skin nodules analyzed by rt-PCR, 31 (70.45 %) were positive while, non of the buffalo samples were positive. Out of 31 positive rt-PCR samples, LSDV was isolated on CAM (n=19; 61.29%) and MDBK cell culture. The virus isolates were confirmed by conventional PCR where 1237 bp product size was successfully amplified. The phylogenetic analysis of LSDV002 gene revealed that three sequenced LSDV isolates were identical to each other and to LSDV isolates from different countries in Africa, Asia, and Europe with 99–100 % identity. ELISA analyses showed seroreactivity of LSDV in Egyptian cattle and buffaloes. In conclusion, the Egyptian water buffalo serves as an accidental non-adapted host for the disease and this point requires more deep investigation. In addition, the current vaccine strategy should be re-evaluated for more coverage and effectiveness.

Identification and Genetic Characterization of Mycoplasma Species Affecting Respiratory System in Egyptian Cattle

Abstract: Mycoplasma species are the main important causative agents causing pneumonia in cattle, there are about 200 species or more. The current work aimed to investigate various mycoplasma spp. isolated from the respiratory tract of cattle by microbial culture, conventional polymerase chain reaction technique, and target gene sequencing with phylogenetic analysis. An initial screening was done to confirm the presence of mycoplasma spp. by culture on PPLO’s agar, digitonin sensitivity and biochemical tests. 129 isolates were characterized by fried egg with depressed center colonies, digitonin sensitive and negative to glucose fermentation and arginine utilization test. Out of 305 samples, 20 samples were selected for amplification by PCR technique using Mycoplasma 16S rRNA primer. Seven samples were positive to Mycoplasma species and gave amplified band at 1013 bp. subsequently, the seven isolates were sequencing. Four sequenced isolates (EGS1, EGL6, EGL1 and EGS2) were closely related to each other and very close to Mycoplasma bovis strains and far from M. leachii 99/014/6. Two sequenced isolates (EGL2 and EGL5) were closely related to each other and very close to Mycoplasma bovirhinis strains and far from M. leachii 99/014/6. One sequenced isolate (EGL3) was very close to Mycoplasma arginini strain and far from M. bovis 99/014/6. From these results, we conclude that conventional culture methods for diagnosis either by isolation and identification of Mycoplasma is a time-consuming method to diagnose mycoplasma infection So, these methods can be replaced by PCR and genome analysis technology.

Seroprevalence, Risk Factors and Molecular Identification of Bovine Leukemia Virus in Egyptian Cattle.

Simple SummaryBovine leukemia virus (BLV) is distributed worldwide and affects dairy cattle causing significant economic losses. This study’s objective was to assess the risk factors associated with BLV infection and identify the Egyptian BLV strain’s genetic diversity. The overall seroprevalence of BLV infection in Egyptian dairy cattle was 18.2%, and the grazing cattle in the losing house system had a higher probability of getting BLV infection. The sequencing and phylogenetic analysis for one Egyptian BLV strain was performed, and the obtained results confirmed the clustering of Egyptian BLV strain into genotype-1.Bovine leukemia virus (BLV) is distributed worldwide and affects dairy cattle causing severe economic losses. The BLV has been serologically reported in Egypt, but few studies have evaluated its associated risk factors and genetic classification. Therefore, this study assessed risk factors associated with BLV infection and identified the genetic diversity of the Egyptian strain. The study was conducted on 500 dairy cattle distributed in four Governorates located in Northern Egypt. Overall, the seroprevalence of BLV infection among Egyptian dairy cattle was 18.2%. The grazing cattle in the losing house system had higher odds for BLV seropositivity, and bad practice such as the use of a single needle or one plastic glove for more than one animal was considered a significant risk factor for BLV infection. Besides, the sequencing and phylogenetic analysis for one Egyptian BLV strain was performed, and the obtained results confirmed the clustering of Egyptian BLV strain into genotype-1. The assessment of associated risk factors for BLV infection and determination of its genetic classification are essential to implement an effective control program.

Effect of polyphenols extracted from Punica granatum and Acacia saligna plants on glutathione S-transferase of the cattle tick Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae).

Ticks are hematophageal ectoparasites that transport major pathogens around the world. Glutathione S-transferases (GST) are involved in resistance to acaricide and redox balancing during the life cycle of the tick. The inhibition of tick GST enzymes by certain phenolic compounds, such as phenolic acids and tannins, can be a promising approach to tick control. The objective of this study was to evaluate the effects of Punica granatum red peel and Acacia saligna leaf extracts on Rhipicephalus (Boophilus) annulatus GST activity in order to reduce the resistance of cattle to acaricide. The results showed that P. granatum ethanol extract (70%) contained the highest total phenol content (350 ± 1.2μM GAE g-1), the highest condensed tannin content (270 ± 1.3μM CE g-1) and the highest hydrolysable tannin content (70 ± 5.0μM TAE g-1). Adult immersion test with a dosage of 100mg ml-1 of A. saligna ethanol extracts had a significant mortality of 50% and 75% after 24h and 96h, respectively (p < 0.01). A simple and reproducible procedure was established to purify the whole R. annulatus GST (wRaGST) while a full-length cDNA of GST was cloned from a cDNA library of the local Egyptian cattle tick R. (B.) annulatus (rRaGST). Aqueous extracts of P. granatum inhibited both wRaGST and rRaGST with values of IC50 = 0.114 and 0.07µgml-1, respectively, compared to A. saligna extracts (IC50 values = 2.08 and 1.35µgml, respectively). These inhibitory effects were attributed to the presence of a high tannin concentration (≥ 80%). HPLC analysis indicated the presence of gallic acid and catechin in both extracts, in addition to the rutin, which was only observed in A. saligna extracts. The addition of a tannin inhibitor, polyethylene glycol, suggested the existence of other phenolic compounds in combination with catechins responsible for inhibiting the activity of these extracts. Non-competitive behaviour of catechins may be helpful in preventing, or at least delaying, the development of chemical acaricide resistancein R. annulatus.