Carcinogenic and non-carcinogenic health risks associated with the consumption of fishes contaminated with heavy metals from Manzala Lake, Egypt.

Manzala Lake was sampled to assess the concentrations and possible ecological risks of heavy metals. The mean heavy metal levels in the muscles of Nile tilapia, Flathead grey mullets and African catfish were 0.01, 0.15 and 0.29 mg/kg, respectively, for mercury; 3.16, 4.25 and 4.74 mg/kg for arsenic; 1.01, 0.87 and 0.95 mg/kg for lead; and 0.05, 0.12 and 0.06 mg/kg for cadmium. The levels of heavy metals exceeded their maximum permissible limits in most samples. The EDIs of some metals were higher than their PTDIs or BMDLs. The THQs and TTHQs from metal intake were >1 for Hg and Cd. In addition, the TCR values of As in all fish species were higher than 1.0 × 10-4 indicating a potential health risks from consumption of fish species which need strict hygienic procedures to prevent fish contamination with heavy metals and ensure that their levels did not exceed the maximum permissible limits.

Evaluation of the Inhibitory Effect of Moringa oleifera Leaves Methanolic Extract against In Vitro Growth of Several Babesia Species and Theileria equi and the In Vivo Growth of Babesia microti.

The current study evaluated the inhibitory effect of Moringa oleifera leaves methanolic extract (MOL) against the in vitro growth of Babesia bovis (B. bovis), B. caballi, B. bigemina, and Theileria equi (T. equi), as well as in vivo growth of B. microti in mice. Active principles of MOL extract were determined using liquid chromatography mass spectrometry (LC-MS). MOL's anti-piroplasm efficacy was assessed both in vitro and in vivo using the SYBR Green I fluorescence assay. Every 96 hours, the hematological parameters, including red blood cell count (RBCs; 104/UL), hemoglobin content (HGB; g/dl), and hematocrit percent (HCT; %), in the treated mice were monitored using a Celltac MEK6450 automated hematological analyzer. LC-MS of MOL revealed that the most abundant polyphenolic catechism found in the MOL extract was isoquercetin and rutin. MOL inhibited B. bovis, B. caballi, B. bigemina, and T. equi in vitro growth in a dose-dependent way, with IC50 values of 45.29 ± 6.14, 19.16 ± 0.45, 137.49 ± 16.07, and 9.29 ± 0.014 µg/ml, respectively. MOL's in vitro antibabesial activity was enhanced when administrated simultaneously with either diminazene aceturate (DA) or MMV665875 compound from malaria box. In mice infected by B. microti, a combination of MOL and a low dose of DA (12.5 mg·kg-1) resulted in a significant (P < 0.05) reduction in B. microti growth. These findings suggest that MOL is an effective herbal anti-piroplasm therapy, especially when combined with a low dosage of either DA or MMV665875.

Prevalence of Multidrug-Resistant Pseudomonas aeruginosa Isolated from Dairy Cattle, Milk, Environment, and Workers' Hands.

Pseudomonas aeruginosa is an opportunistic pathogen causing severe infection in animals and humans. This study aimed to determine the ecological distribution and prevalence of multidrug-resistant (MDR) P. aeruginosa isolated from dairy cattle, the environment, and workers' hand swabs. Samples (n = 440) were collected from farms and households (n = 3, each). Rectal swabs, udder skin swabs, milk, workers' hand swabs, feed, water, water sources, and beddings were collected. Samples were subjected to the bacterial identification of P. aeruginosa via 16S rRNA. Antimicrobial resistance (AMR) was detected either phenotypically using an antibiotic susceptibility test or genotypically with AMR resistance genes (ARGs) such as drfA, sul1, and ermB. P. aeruginosa was detected on dairy farms and households (10.3-57.5%, respectively), with an average of 23.2%. The resistance of dairy farm strains was observed against sulfamethoxazole, imipenem, cefepime, piperacillin-tazobactam, and gentamycin (100%, 72.7%, 72.7%, 68.8%, and 63.3%, respectively). Meanwhile, the resistance of household strains was observed against sulfamethoxazole, imipenem, amoxicillin, gentamicin, cefepime, and erythromycin by 91.3%, 82.6%, 75.4%, 75.4%, 68.1%, and 63.8%, respectively. The susceptibility of farm strains was detected against norfloxacin, ciprofloxacin, and levofloxacin (90.9%, 84.8%, and 72.7%, respectively). Meanwhile, the susceptibility of household strains was detected against ciprofloxacin, amikacin, and norfloxacin (100%, 84.1%, and 72.5%, respectively). About 81.4% of P. aeruginosa strains were MDR. ARGs (drfA, sul1, and ermB) were detected in farm strains (48.5%, 72.7%, and 24.4%, respectively) and household strains (50.7%, 72.5%, and 47.8%, respectively). Almost all P. aeruginosa had MAR over 0.2, indicating repeated application of antibiotics. P. aeruginosa prevalence was fivefold higher in households than on farms. MDR strains were higher amongst household strains than farm strains.

Antiparasitic activity of FLLL-32 against four Babesia species, B. bovis, B. bigemina, B. divergens and B. caballi, and one Theileria species, Theileria equi in vitro, and Babesia microti in mice.

Introduction: FLLL-32, a synthetic analog of curcumin, is a potent inhibitor of STAT3's constitutive activation in a variety of cancer cells, and its anticancer properties have been demonstrated both in vitro and in vivo. It is also suggested that it might have other pharmacological activities including activity against different parasites. Aim: This study therefore investigated the in vitro antiparasitic activity of FLLL-32 against four pathogenic Babesia species, B. bovis, B. bigemina, B. divergens, and B. caballi, and one Theileria species, Theileria equi. In vivo anti-Babesia microti activity of FLLL-32 was also evaluated in mice. Methods: The FLLL-32, in the growth inhibition assay with a concentration range (0.005-50 µM), was tested for it's activity against these pathogens. The reverse transcription PCR (RT-PCR) assay was used to evaluate the possible effects of FLLL-32 treatment on the mRNA transcription of the target B. bovis genes including S-adenosylhomocysteine hydrolase and histone deacetylase. Results: The in vitro growth of B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi was significantly inhibited in a dose-dependent manner (in all cases, p < 0.05). FLLL-32 exhibits the highest inhibitory effects on B. bovis growth in vitro, and it's IC50 value against this species was 9.57 µM. The RT-PCR results showed that FLLL-32 inhibited the transcription of the B. bovis S-adenosylhomocysteine hydrolase gene. In vivo, the FLLL-32 showed significant inhibition (p < 0.05) of B. microti parasitemia in infected mice with results comparable to that of diminazene aceturate. Parasitemia level in B. microti-infected mice treated with FLLL-32 from day 12 post infection (pi) was reduced to reach zero level at day 16 pi when compared to the infected non-treated mice. Conclusion: The present study demonstrated the antibabesial properties of FLLL-32 and suggested it's usage in the treatment of babesiosis especially when utilized in combination therapy with other antibabesial drugs.

Seroprevalence of Rift Valley Fever virus in one-humped camels (Camelus dromedaries) in Egypt.

Rift Valley fever (RVF) is a mosquito-borne viral disease that affects a variety of domestic animals, including cattle, sheep, goats, and camels, and has zoonotic potential. Although the rift valley fever virus (RVFV) is usually asymptomatic in camels, it can induce abortion in some pregnant animals. In the current study, a serosurvey was carried out to investigate the prevalence of RVFV antibodies and related risk factors in camels from four Egyptian governorates. A total of 400 serum samples were examined for anti-RVFV antibodies using a competitive enzyme-linked immunosorbent assay (c-ELISA). The results revealed that the overall prevalence of RVF among examined camels was 21.5% and the disease was more prevalent in Kafr ElSheikh governorate in Nile Delta of Egypt. In addition, the age group of camels with more than 5 years (OR=4.49, 95%CI: 1.39-14.49), the female sex (OR=3.38, 95%CI: 1.51-7.58), the emaciated animals (OR=1.52, 95%CI: 0.86-2.66), the summer season's infection (OR=5.98, 95%CI: 1.79-19.93), the presence of mosquitoes (OR= 2.88, 95%CI: 1.39-5.95), and the absence of mosquitoes control (OR=3.97, 95%CI: 2.09-7.57) were identified as risk factors for RVFV infection. The results of this study support knowledge on the risk factors for RVFV infection and demonstrate that camels raising in Egypt have RVFV antibodies. Quarantine measures or vaccination program should be implemented to reduce the likelihood of RVFV introduction, dissemination among susceptible animals, and ultimately transmission to humans.

Effect of nanostructure lipid carrier of methylene blue and monoterpenes as enzymes inhibitor for Culex pipiens.

Solid lipid nanoparticles second generation, nanostructure lipid carrier (NLC), is one of the most important biodegradable nanoparticles. Nanostructure Lipid carrier (NLC) was used to encapsulate methylene blue (MB) dye, carvacrol and citronellal and their efficacy as insecticidal against Culex pipiens (Cx. pipiens) were distinguished. The prepared nanoformulation revealed very good physicochemical properties, especially the homogeneity of the particle size. Transmission electron microscope showed spherical shaped nanoparticles within range less than 200 nm. The prepared NLC-MB-MT system showed a very competitive insecticidal activity and high virulence against the mosquito larvae with higher mortality rate of LC50 of 0.141 µl/mL, in addition to high level of Oxidative stress parameters obtained through all the tested enzymes including hydrogen peroxide (4.8 ppm), protein carbonyl amount (0.12 OD/mg protein), ascorbic acid (0.15 mg) and Superoxide dismutase (SOD) showed strong increasing (0.09 OD/mg protein/min) at 6 µg/mL, respectively. Whereas paradoxical results of the oxidative stress enzymes were obtained from different concentration of nanoformulation that introduce a convenient reason for their potential insecticidal effect. The cytotoxic effect of NLC-MB-MT was evaluated using WI38 human lung cell lines, the LC50 was 6.4 mg/mL. The low cytotoxic reactivity towards the tested cell line makes the NLC-MB-MT nanoformulation has its promising insecticidal efficacy. Molecular docking study for each component were done against acetylcholine esterase protein and accepted binding modes achieved by the three compounds.

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.

Evaluating the inhibitory effect of resveratrol on the multiplication of several Babesia species and Theileria equi on in vitro cultures, and Babesia microti in mice.

Introduction: Histone post-translational modification is one of the most studied factors influencing epigenetic regulation of protozoan parasite gene expression, which is mediated by histone deacetylases (KDACs) and acetyltransferases (KATs). Objective and methods: The present study investigated the role of resveratrol (RVT) as an activator of histone deacetylases in the control of various pathogenic Babesia sp. and Theileria equi in vitro, as well as B. microti infected mice in vivo using fluorescence assay. Its role in mitigating the side effects associated with the widely used antibabesial drugs diminazene aceturate (DA) and azithromycin (AZM) has also been investigated. Results: The in vitro growth of B. bovis, B. bigemina, B. divergens, B. caballi and Theileria equi (T. equi) was significantly inhibited (P < 0.05) by RVT treatments. The estimated IC50 values revealed that RVT has the greatest inhibitory effects on B. bovis growth in vitro, with an IC50 value of 29.51 ± 2.46 µM. Reverse transcription PCR assay showed that such inhibitory activity might be attributed to resveratrol's stimulatory effect on B. bovis KDAC3 (BbKADC3) as well as its inhibitory effect on BbKATS. RVT causes a significant decrease (P < 0.05) in cardiac troponin T (cTnT) levels in heart tissue of B. microti- infected mice, thereby indicating that RVT may play a part in reducing the cardiotoxic effects of AZM. Resveratrol showed an additive effect with imidocarb dipropionate in vivo. Treatment of B. microti-infected mice with a combined 5 mg/kg RVT and 8.5 mg/kg ID resulted in an 81.55% inhibition at day 10 postinoculation (peak of parasitemia). Conclusion: Our data show that RVT is a promising antibabesial pharmacological candidate with therapeutic activities that could overcome the side effects of the currently used anti-Babesia medications.

Magnesium Supplementation Alleviates the Toxic Effects of Silica Nanoparticles on the Kidneys, Liver, and Adrenal Glands in Rats.

Concerns regarding the possible hazards to human health have been raised by the growing usage of silica nanoparticles (SiNPs) in a variety of applications, including industrial, agricultural, and medical applications. This in vivo subchronic study was conducted to assess the following: (1) the toxicity of orally administered SiNPs on the liver, kidneys, and adrenal glands; (2) the relationship between SiNPs exposure and oxidative stress; and (3) the role of magnesium in mitigating these toxic effects. A total of 24 Sprague Dawley male adult rats were divided equally into four groups, as follows: control group, magnesium (Mg) group (50 mg/kg/d), SiNPs group (100 mg/kg/d), and SiNPs+ Mg group. Rats were treated with SiNPs by oral gavage for 90 days. The liver transaminases, serum creatinine, and cortisol levels were evaluated. The tissue malondialdehyde (MDA) and reduced glutathione (GSH) levels were measured. Additionally, the weight of the organs and the histopathological changes were examined. Our results demonstrated that SiNPs exposure caused increased weight in the kidneys and adrenal glands. Exposure to SiNPs was also associated with significant alterations in liver transaminases, serum creatinine, cortisol, MDA, and GSH. Additionally, histopathological changes were significantly reported in the liver, kidneys, and adrenal glands of SiNPs-treated rats. Notably, when we compared the control group with the treated groups with SiNPs and Mg, the results revealed that magnesium could mitigate SiNPs-induced biochemical and histopathologic changes, confirming its effective role as an antioxidant that reduced the accumulation of SiNPs in tissues, and that it returns the levels of liver transaminases, serum creatinine, cortisol, MDA, and GSH to almost normal values.

Impact of chitosan administration on titanium dioxide nanoparticles induced testicular dysfunction.

The potential reproductive toxic effects of oral TiO2 NPs in adult male rats as well as the possible alleviation of chitosan administration was investigated. Animals were allocated to four groups; the first group received deionized water and was assigned as a control group. In the second group, rats received chitosan at a dose of 5 mg/kg BW/day. The third group was designed for administration of TiO2 NPs at a dose of 150 mg/kg BW/day (1/80 LD50). Rats in the fourth group received both TiO2 NPs and chitosan. After 14 days, TiO2 NPs induced testicular lipid peroxidation as well as oxidative stress. Nano-titanium significantly upregulated genes that encode apoptosis and inflammation in testicular tissue. Moreover, it induced histological alteration in the testicular structure with impairment in spermatogenesis via reduction of PCNA immune-staining. Chitosan administration significantly improved the activities of testicular GPx, SOD, and CAT enzymes. In addition, it significantly down-regulated the relative expressions of pro-apoptotic and pro-inflammatory testicular genes. Chitosan was able to improve the testicular architecture as well as spermatogenesis. The current study revealed the capability of chitosan to ameliorate nano-titanium induced testicular toxicity. Thus, attention should be given to the extensive consumption of nano-titanium particles.

Ecological Distribution of Virulent Multidrug-Resistant Staphylococcus aureus in Livestock, Environment, and Dairy Products.

Staphylococcus aureus is one of the most common causes of mastitis, leading to severe economic losses in the dairy industry. It is also zoonotic, with potential risks to public health. This study aimed to detect the occurrence of S. aureus-resistant strains isolated from cattle, buffalo, their environment, milk and dairy products; and to investigate the extent of animal, ecological, and food contamination by methicillin-resistant (MRSA) or enterotoxigenic S. aureus. Samples (n = 350) were collected from four animal (two cattle and two buffalo) farms, i.e., their environment. Thirty Karish cheese samples were collected from 10 markets in Mansoura, Egypt. S. aureus was detected in 17.9%, 17.6%, and 16.7% of samples collected from cattle, buffalo and Karish cheese, respectively. About 19% of isolated S. aureus strains carried the mecA gene. The distribution of the mecA gene was high in isolates from Karish cheese (60%), followed by samples collected from buffalo (16.2%) and cattle (16%). More than 34% of isolated S. aureus strains were enterotoxigenic, and the presence of enterotoxin genes was higher in isolates from Karish cheese (80%) than those from cattle (48%) and buffalo (18.9%). The most predominant enterotoxin gene among isolated S. aureus strains was the sea gene (26.9%), followed by sec (4.5%) and sed (3%) genes. Isolated strains were resistant to clindamycin (100%), kanamycin (97%), nalidixic acid (86.6%), cefotaxime (73.1%) sulphamethazole-trimethoprim (65.6%). Meanwhile, 95.5%, 94%, 86.6% and 77.7% of S. aureus strains were sensitive to ciprofloxacin, amikacin, imipenem and both cefoxitin and gentamycin, respectively. In conclusion, the presence of enterotoxigenic- and methicillin-resistant S. aureus strains in animals, their environment, and dairy products represents a public health concern, particularly in small-scale dairy farms in Egypt. To reduce the risk of infection of livestock and humans with resistant strains, strict regulations and guidelines for antimicrobial use in such a system are urgently required.

Molecular survey and phylogenetic analysis of Babesia vogeli in dogs.

Canine babesiosis is a life-threatening haemoparasitic disease in dogs that is prevalent worldwide. In this study, the prevalence of Babesia vogeli (B. vogeli) was investigated in dogs from Egypt by using Polymerase Chain Reaction (PCR) assay, and associated risk factors were evaluated. In addition, phylogenetic position of B. vogeli Egyptian isolate was determined by sequencing. A total of 275 blood samples were taken from dogs located in four governorates belonging to the north of Egypt. Samples were examined by PCR targeting the B. vogeli 18S rRNA gene and this species was also confirmed by sequencing. Overall, the prevalence of B. vogeli was 5.1% among the studied dogs and the highest prevalence rate was found in the Giza governorate. Univariate logistic regression was used to evaluate each variable individually. The results revealed a significant association between the prevalence of B. vogeli infection and whether or not dogs were infested with ticks and the type of floor used in dog shelters. Additionally, tick infestation (OR 6.1, 95% CI 1.2-31.4), and living in shelters with soil floors (OR 3.8, 95% CI 0.8-17.8) were identified as potential risk factors for B. vogeli infection. Phylogenetic analysis was performed using B. vogeli 18S rRNA partial sequences with the hypervariable V4 region from GenBank. The Egyptian isolate was assigned to second sub-cluster with B. vogeli isolates from Japan, Venezuela and Paraguay within the B. vogeli/B. canis cluster. The present data will be useful to improve the understanding of canine babesiosis epidemiology and ways to control the disease in companion dogs.

Detection of Equid Alphaherpesvirus 1 from Arabian Horses with different clinical presentations between 2016-2019 in Egypt.

Equid alphaherpesvirus 1 (EHV-1) is an important virus causing pathological disorders in horses. This highly contagious pathogen causes persistent outbreaks of upper respiratory tract infection, ocular affections, abortion, and neurological disorders with high mortality in Arabian horses in Egypt. The quick and accurate diagnosis is important to broaden our understanding about EHV-1 in the field, and to implicate stronger preventive, and control measures. Sixty-six Arabian horses from Cairo and Giza governorates were sampled from respiratory, abortigenic and neurological outbreaks over a period of 4 years. EHV-1 was diagnosed in these cases by immunohistochemistry using monoclonal antibody against EHV-1 glycoprotein B and molecular detection using gB, ORF33 specific real-time PCR. EHV-1 was detected in 25 cases, mostly from abortigenic outbreaks (14 abortions, 3 stillbirths, and two early neonatal deaths), in addition to 5 respiratory affections and single EHV-1 myeloencephalopathy. Molecular characterization revealed that the ORF33 sequences from this study were almost identical and closely related to the European EHV-1 strains. Furthermore, no difference in the amino acid sequences compared to previously published EHV-1 sequences from Egypt. The data in this study provides some insights about the prevalance of EHV-1 infection in Arabian horses, discusses EHV-1 diagnostic approaches, highlights the importance of accurate diagnosis and the importance of pregnant mare vaccination, and adds to the previous knowledge about EHV-1 in Egypt which may help in better controlling EHV-1 infections in the future.

Assessment of Seroprevalence and Associated Risk Factors for Anaplasmosis in Camelus dromedarius.

(1) Background: Anaplasmosis is an infectious disease in camels caused by an obligate intracellular bacterium that is transmitted by ticks. (2) Methods: A cross-sectional study was conducted during 2020 to study the seroprevalence of Anaplasma spp. among Camelus dromedarius in three governorates in Egypt and assess the associated risk factors. Serum samples from 365 camels were examined by a competitive enzyme-linked immunosorbent assay (cELISA) test. (3) Results: Overall, the seroprevalence of anaplasmosis among camels was 18.6%. Multivariable logistic regression was performed, and it was discovered that tick infestation, application of acaricides, grooming practice and body condition were potential risk factors for Anaplasma spp. infection (odds ratio > 1) in dromedary camels. In contrast, the locality in which the camels lived and their age were not significant effects with regard to the occurrence of anaplasmosis. (4) Conclusions: The current findings suggest that improvement of protective measures to limit the effects of the identified risk factors can help to reduce the spread of anaplasmosis among camels in Egypt.

Cross-sectional survey on Mycobacterium avium Subsp. paratuberculosis in Dromedary Camels: Seroprevalence and risk factors.

Johne's disease is a chronic disease with great concern in ruminants and caused by Mycobacterium avium subsp. paratuberculosis (MAP). A cross-sectional study was conducted from February 2019 to January 2020 to estimate the prevalence of MAP infection among camels which are kept in three governorates in Nile Delta of Egypt. A total of 440 serum samples were examined by ELISA for detection of MAP antibodies. The multivariable logistic regression model was performed to determine the associated risk factors for MAP infection in examined camels. Overall, the seroprevalence of MAP infection was found to be 7.5% among examined camels. The multivariable logistic regression model was performed to determine the associated risk factors for MAP infection in examined camels. The main findings revealed that the risk of getting MAP infection increased among elder camels (>10 years old) with signs of diarrhea, having communal water source and in camels grazing in the same pasture (odds ratio >1). However, geographic location, sex and contact with cattle had not significant impact regarding to seroprevalence of MAP infection in camels. The present findings confirm presence of MAP among camels which is a potential risk factor for contamination of environment and spreading of infection. Therefore, further studies for detection of infected animals in early stage are needed beside the estimated risk factors in this study to build an efficient control program.

Immunomodulatory Effect of Vitamin C on Proinflammatory Cytokines Production in Ossimi Lambs (Ovis aries) with Pneumonic Pasteurellosis.

In this study, we have investigated the impact of vitamin C on the production of pro-inflammatory cytokines (interleukin 1 ß (IL-1 ß), interleukin 6 (IL-6), interleukin 12p40 (IL-12p40), interferon gamma (IFNγ), and tumor necrosis factor alpha (TNF-α)) in lambs naturally infected by pneumonic pasteurellosis. Of 37 lambs, 18 lambs were identified to have pneumonic pasteurellosis and randomly allocated into two equal groups. Single subcutaneous dose of tulathromycine alone (2.5 mg kg-1) or tulathromycine combined with vitamin C (3 gm kg-1) were administrated to the diseased lambs. The serum levels of IL-1ß, IL-6, IFN-γ, and TNF-α were returned to the normal levels in pneumonic lambs treated with the combination therapy. The obtained results indicate the selective influences of vitamin C on pro-inflammatory cytokines production in sera of lambs with pneumonic pasteurellosis and highlights the value of vitamin C as a potential anti-inflammatory drug and ideal immunomodulatory agent.

Identification and Characterization of P0 Protein as a Vaccine Candidate Against Babesia divergens, Blood Parasite of Veterinary and Zoonotic Importance.

The molecular identification and antigenic characterization of P0 protein in Babesia divergens, a blood parasite of veterinary and zoonotic importance, were carried out in this study for use in developing subunit vaccines against B. divergens infection. Recombinant protein encoding P0 (BdP0) was developed in Escherichia coli, and its antiserum was generated in mice for further molecular characterization. Anti-rBdP0 serum had a specific interaction with the corresponding legitimate B. divergens protein, as confirmed by Western blotting and indirect fluorescent antibody tests. ELISA was used to assess the immunogenicity of BdP0 in a group of 68 bovine field samples, and significant immunological reactivity was found in 19 and 20 positive samples of rBdp0 and B. divergens lysate, respectively. The in vitro growth of B. divergens cultures treated with anti-rBdP0 serum was significantly inhibited (p < 0.05). Furthermore, after 6 h of incubation with 2 mg/ml anti-rBdP0 serum, the ability of pre-incubated free merozoites to invade bovine erythrocytes was reduced by 59.88%. The obtained data suggest the possible use of rBdP0 as diagnostic antigen and may serve as a vaccine candidate against babesiosis caused by B. divergens either in animal or human.