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.

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.

Seroprevalence and risk factors analysis of Neospora caninum and Toxoplasma gondii in cattle of Beheira, Egypt.

Introduction: Neospora caninum and Toxoplasma gondii are closely related obligate intracellular protozoan parasites. They are considered to be the major causes of infectious abortions and congenital abnormalities in livestock worldwide resulting in huge economic losses. Currently, there are no reports on the prevalence of neosporosis or toxoplasmosis in cattle in Beheira, Egypt's most important region for cattle industry. Methods: The current study investigated the presence of anti-N. caninum and anti-T. gondii antibodies in apparent healthy cattle from eight localities representing the whole area of Beheira. A total of 358 plasma samples were randomly collected from 6 dairy and 10 beef farms and analyzed by commercially available ELISAs. Production type (dairy versus beef), sex (female vs male), age (< 3 years, 3-5, and > 5 years old), breed (mixed vs Holstein vs Colombian Zebu), and location (various locations) were assessed as risk factors for N. caninum and T. gondii infections. Results and discussion: Of the samples, 88 (24.6%) and 19 (5.3%) were positive for anti-N. caninum and anti-T. gondii antibodies, respectively, and mixed infection was detected in 7. Of the 16 herds, 6 dairy and 7 beef herds were positive for antibodies to N. caninum. Antibodies to T. gondii were detected in 4, and 5 of dairy and beef herds, respectively. Production type (dairy) and, therewith, sex (female), age (aged over 5 years), and location were considered as risk factors for N. caninum infection. No factors statistically associated with T. gondii infection were identified. Overall, this study provided the first serological detection of N. caninum and T. gondii infections in cattle from Beheira, demonstrating the endemicity of both parasites in the main cattle rearing region of Egypt. This study also confirmed earlier reports of N. caninum being more present in dairy cattle than in beef cattle. Routine monitoring of N. caninum and T. gondii infections and the implementation of control strategies are urgently needed.

A Newly Emerging Serotype A Strain in Foot-and-Mouth Disease Virus with Higher Severity and Mortality in Buffalo than in Cattle Calves in North Egypt.

A severe foot-and-mouth disease (FMD) epidemic struck several Egyptian provinces recently, causing significant losses among animals even in vaccinated farms. This study indicated the existence of the newly emerging foot-and-mouth disease virus (FMDV) and first investigated its effect on the Egyptian water buffalo (Bubalus bubalis) and cattle calves in the Beheira province, north Egypt. Twenty tongue epithelial samples from diseased calves in five infected farms were randomly collected, prepared, and propagated using baby hamster kidney-21 (BHK-21) cells. Whole genomic RNA was extracted from the cells of the third passage. A FMDV genome was detected and serotyped using one-step reverse transcription polymerase chain reactions (RT-PCRs). Nucleotide sequencing of the purified serotype-specific PCR bands was performed, and a maximum likelihood phylogenetic tree based on 600 base pairs of VP1 was constructed. The results identified FMDV, serotype A in all infected samples, whereas the serotypes O and SAT2 were negative. The obtained 20 sequences were identical to each other and similar to the newly reported strain in Egypt that belongs to the Europe-South America (Euro-SA) topotype. The epidemiological and clinical parameters associated with such a strain were fully recorded by veterinarians and analyzed in a single infected farm including 70 cattle and buffalo calves. It caused higher peracute mortalities in buffalo (25.7%; 95% CI: 13-43) than in cattle (8.6%; 95% CI: 2-24) calves. Severe clinical signs such as dullness, hypothermia, bradycardia, and cardiac arrhythmia were common to both except in fatal cases, whereas hyperthermia and respiratory signs were prevalent in cattle calves. In conclusion, we first characterized the newly emerging FMDV in the calves of Beheira as more fatal and severe in buffalo than in cattle calves.

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.

BoLA-DRB3 Polymorphism Associated with Bovine Leukemia Virus Infection and Proviral Load in Holstein Cattle in Egypt.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, the most prevalent neoplastic disease of cattle worldwide. The immune response to BLV and disease susceptibility and resistance in cattle are strongly correlated with the bovine leukocyte antigen (BoLA)-DRB3 allelic polymorphism. BLV infection continues to spread in Egypt, in part because the relationships between BLV infection, proviral load in Egypt, and BoLA-DRB3 polymorphism are unknown. Here, we identified 18 previously reported alleles in 121 Holstein cows using a polymerase chain reaction sequence-based typing method. Furthermore, BoLA-DRB3 gene polymorphisms in these animals were investigated for their influence on viral infection. BoLA-DRB3*015:01 and BoLA-DRB3*010:01 were identified as susceptible and resistant alleles, respectively, for BLV infection in the tested Holsteins. In addition, BoLA-DRB3*012:01 was associated with low PVL in previous reports but high PVL in Holstein cattle in Egypt. This study is the first to demonstrate that the BoLA-DRB3 polymorphism confers resistance and susceptibility to PVL and infections of BLV in Holstein cattle in Egypt. Our results can be useful for the disease control and eradication of BLV through genetic selection.

High relatedness of bioinformatic data and realistic experimental works on the potentials of Fasciola hepatica and F. gigantica cathepsin L1 as a diagnostic and vaccine antigen.

Introduction: Fascioliasis is a parasitic foodborne disease caused by the liver flukes, Fasciola hepatica and F. gigantica. Such parasites cause serious illness in numerous domestic animals and also in humans. Following infection, the parasite secretes a variety of molecules that immediately interact with the host immunity to establish successful infection. These molecules include cathepsin L peptidase 1 (CatL1); the highly investigated diagnostic and vaccine antigens using various animal models. However, a few studies have analyzed the potentials of FhCatL1 as a diagnostic or vaccine antigen using bioinformatic tools and much less for FgCatL1. The present study provides inclusive and exclusive information on the physico-chemical, antigenic and immunogenic properties of F. hepatica cathepsin L1 (FhCatL1) protein using multiple bioinformatic analysis tools and several online web servers. Also, the validation of our employed available online servers was conducted against a huge collection of previously published studies focusing on the properties of FhCatL1as a diagnostic and vaccine antigen. Methods: For this purpose, the secondary, tertiary, and quaternary structure of FhCatL1 protein were also predicted and analyzed using the SWISS-MODEL server. Validation of the modeled structures was performed by Ramachandran plots. The antigenic epitopes of the protein were predicted by IEDB server. Results and discussion: Our findings revealed the low similarity of FhCatL1 with mammalian CatL1, lacking signal peptides or transmembrane domain, and the presence of 33 phosphorylation sites. Also, the containment of FhCatL1 for many topological, physico-chemical, immunological properties that favored its function of solubility and interaction with the immune components were reported. In addition, the earlier worldwide reports documented the high efficacy of FhCatL1 as a diagnostic and vaccine antigen in different animals. Altogether, FhCatL1 is considered an excellent candidate for using in commercialized diagnostic assays or vaccine products against fascioliasis in different animal species. Our assessment also included FgCatL1 and reported very similar findings and outputs to those of FhCatL1.

Toxoplasma gondii and Neospora caninum Antibodies in Dogs and Cats from Egypt and Risk Factor Analysis.

BACKGROUND: Toxoplasma gondii and Neospora caninum are major protozoan parasites of worldwide distribution and significance in veterinary medicine and, for T. gondii, in public health. Cats and dogs, as final hosts for T. gondii and N. caninum, respectively, have a key function in environmental contamination with oocysts and, thus, in parasite transmission. Very little is known about the prevalence of T. gondii infections in dogs and cats in Egypt, and even less about the prevalence of N. caninum in the same hosts. METHODS: In the current study, 223 serum samples of both dogs (n = 172) and cats (n = 51) were investigated for specific antibodies to T. gondii and N. caninum using commercially available ELISAs. A risk factor analysis was conducted to identify factors associated with seropositivity. RESULTS & DISCUSSION: Exposure to T. gondii was reported in 23.3% of the dogs and in 9.8% of the cats, respectively. In addition, N. caninum-specific antibodies were recorded in 5.8% of dogs and in 3.4% of cats. A mixed infection was found in two dogs (1.2%) and in one cat (2%). Antibodies to T. gondii in dogs were significantly more frequent in dogs aged 3 years or more and in male German Shepherds. As this breed is often used as watchdogs and was the most sampled breed in Alexandria governorate, the purpose "watchdog" (compared to "stray" or "companion"), the male sex, and the governorate "Alexandria" also had a significantly higher seroprevalence for T. gondii. No factors associated with antibodies to N. caninum could be identified in dogs, and no significant factors were determined in cats for either T. gondii or N. caninum infection. Our study substantially adds to the knowledge of T. gondii infection in dogs and cats and presents data on N. caninum infection in cats for the first and in dogs in Egypt for the second time.

Detection and Molecular Characterization of Bovine Leukemia Virus in Egyptian Dairy Cattle.

Bovine leukemia virus (BLV) causes enzootic bovine leukosis (EBL), the most common neoplastic disease in cattle worldwide. The first EBL outbreak in Egypt was reported in 1997. To date, there are few studies regarding BLV diagnosis using only serological detection and no studies investigating the distribution of BLV provirus, which is the retroviral genome integrated into the host genome, in Egypt. The genetic characteristics of Egyptian BLV strains are also unknown. Therefore, we aimed to detect BLV provirus and determine BLV genetic variability among dairy cattle in Egypt. We collected 270 blood samples of dairy cattle from 24 farms located in five provinces in Egypt. Out of the 270 samples, 58 (21.5%) were positive for BLV provirus. Phylogenetic analysis based on 18 420-bp selected sequences out of 50 isolates of the BLV env-gp51 gene demonstrated that Egyptian BLV isolates were clustered into genotype-1 and-4, among 11 genotypes detected worldwide. Furthermore, phylogenetic analysis and alignment of the 501-bp sequence of the env-gp51 gene revealed that at least six genetically different strains are present in Egypt. Genotype-1 isolates comprised four different strains (G1-a, G1-b, G1-c, and G1-d) and genotype-4 isolates included two different strains (G4-x and G4-y). Moreover, in one farm with 100% infection rate, we identified three isolates of G1-a strain, 35 isolates of G4-x strain, and two isolates of G4-y strain. Overall, this study provides the new report on molecular prevalence of BLV in Egypt and records the coexistence of BLV genotype-1 and-4 in Egyptian cattle.

Detection and molecular characterization of bovine leukemia virus in beef cattle presented for slaughter in Egypt.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, the most common neoplastic disease of cattle worldwide and a serious problem for the cattle industry. Previous studies have shown the molecular prevalence of BLV and the coexistence of BLV genotype-1 and -4 in Egyptian dairy cattle; however, the molecular characteristics of BLV in Egyptian beef cattle are unknown. Therefore, we collected blood samples of 168 beef cattle from slaughterhouses in three governorates in Egypt. Based on BLV-CoCoMo-qPCR-2 targeting long terminal repeats and nested PCR targeting the env-gp51 gene, the BLV provirus infection rates were found to be 47/168 (28.0%) and 42/168 (25.0%), respectively. Phylogenetic analysis based on 501 bp of the BLV env-gp51 gene from 42 BLV isolates revealed that at least six distinctive strains (b, e, f, g, x, and z) were prevalent in cattle across the examined regions. Furthermore, phylogenetic analysis of the 420 bp sequence of the BLV env-gp51 region of the six strains against 11 known genotypes showed that the strains b, e, f, and g were clustered into genotype-1, and strains x and z were clustered into genotype-4. Our results also indicated that strains b and x exist in both dairy and beef cattle in Egypt. The present study is the first to detect and genotype BLV among beef cattle in Egypt.