Efficacy of probiotic Enterococcus faecium in combination with diclazuril against coccidiosis in experimentally infected broilers.

AIMS: The study was conducted to investigate the combination of a probiotic strain of Enterococcus faecium and diclazuril to control coccidiosis in broilers. METHODS AND RESULTS: A total of 240 one-day-old female broiler chicks were divided into eight groups (30 chicks per group): prophylactic groups (G1, G2 and G3) and therapeutic groups (G4, G5 and G6) and two control groups (untreated infected, G7 and untreated uninfected, G8 controls). In the prophylactic approach, diclazuril alone (G1), probiotic alone (G2) or a mixture of both probiotic and diclazuril (G3) was orally administered to the chicks via drinking water 10 days prior to the infection. However, in the therapeutic approach, G4, G5 and G6 birds were administered diclazuril alone, probiotic alone and diclazuril+probiotic mix, respectively, in drinking water for five consecutive days after the appearance of clinical signs of coccidiosis. Birds of both approaches and G7 were experimentally infected with 25 × 103 Eimeria-sporulated oocysts. Chicks in G3 showed the highest weight gain, the lowest lesion score, a low oocyst count and mortality rate among the challenged groups. Moderate lesion scores and oocyst counts were observed in chickens administered probiotics prophylactically. In the therapeutic approach, broilers in G6 but not G5 displayed a decreased mortality rate and lesion score in comparison to those in G7 and G8. However, the result of the probiotic-treated group was not significantly different from that in the untreated infected control group. CONCLUSION: The probiotic supplementation as a prophylactic approach can decrease the adverse effects of eimerian infection. In addition, the probiotic and diclazuril mix achieved a considerable improvement in the growth performance. Therefore, probiotic plus diclazuril combination achieved a synergistic effect. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigation into the synergism/antagonism between a probiotic and diclazuril as anticoccidial agent and the difference in the timing of administration.

Modulation of murine intestinal immunity by Moringa oleifera extract in experimental hymenolepiasis nana.

The potential therapeutic value of Moringa oleifera extract (MOE), due to its anti-inflammatory and anti-oxidant effects, has been reported previously. In this study, Hymenolepis nana antigen (HNA) in combination with MOE was used in immunization against H. nana infection. Adult worm and egg counts were taken, while histological changes in the intestine were observed. Mucosal mast (MMCs) and goblet cells (GCs) were stained with specific stains, while serum and intestinal IgA were assayed using enzyme-linked immunosorbent assay (ELISA). Reduced glutathione (GSH) and lipid peroxidation (thiobarbituric acid reactive substances, TBARS) were assayed. Real-time polymerase chain reaction (PCR) was used for detection of mRNA expression in ileum tissue. The results demonstrated an improvement in the architecture of intestinal villi, decreased inducible nitric oxide synthase (iNOs) and TBARS, and increased GSH in HNA, MOE and MOE + HNA groups. In the same groups, an increase in GCs, mucin 2 (MUC2), interleukins (IL)-4, -5 and -9, and stem cell factor (SCF) versus a decrease in both interferon-gamma (IFN-γ) and transforming growth factor (TGF-ß) expression appeared. HNA and MOE + HNA increased serum and intestinal IgA, respectively. MOE decreased MMCs and achieved the highest reductions in both adult worms and eggs. In conclusion, MOE could achieve protection against H. nana infections through decreased TGF-ß, IFN-γ and MMC counts versus increased GC counts, T-helper cell type 2 (Th2) cytokines and IgA level.

Fasciola hepatica infections in cattle and the freshwater snail Galba truncatula from Dakhla Oasis, Egypt.

Infection by Fasciola species was investigated in seven districts of Dakhla Oasis, Egypt, through abattoir inspection of cattle livers for adult worms and sedimentation of faecal samples from local cattle to detect Fasciola eggs. In addition, lymnaeid snails collected from the study area were examined microscopically for developmental stages of Fasciola spp. Abattoir inspection revealed that 51 out of 458 cattle livers (11.1%) contained adult flukes, which were identified morphologically as Fasciola hepatica. Examination of the cattle faecal samples revealed that 142 out of 503 (28.2%) contained Fasciola eggs. The collected snails, identified as Galba truncatula and Radix natalensis, showed larval stages of Fasciola in 71 out of 731 (9.7%) G. truncatula, while R. natalensis showed no infection. Specific duplex polymerase chain reaction (PCR) targeting the mitochondrial cox1 gene of F. hepatica and Fasciola gigantica was carried out on DNA extracted from pooled infected snails and adult worms. The F. hepatica size amplicon (1031 bp) was obtained from both the infected G. truncatula and the adult worms isolated from cattle livers from different districts. The amplicon sequences were identical to the published sequences of F. hepatica mitochondrial cox1 gene. In conclusion, the zoonotic importance of Fasciola infection and appropriate hygienic measures must be taken into consideration in Dakhla Oasis, Egypt.

Molecular detection of Rhipicephalus (Boophilus) annulatus resistance against deltamethrin in middle Egypt.

This study was designed to investigate the susceptibility/resistance of Rhipicephalus (Boophilus) annulatus to commercial deltamethrin 5%. The work was based on in vitro bioassays; adult immersion test (AIT) and larval packet test (LPT). Fully engorged female ticks were collected from 11 locations in Beni-Suef Governorate, Egypt (Alfashin, Eminelaros, Ehnasia, Smusta, Belifia, Aldiabia, Alhalabia, Beshnna, Alkom, Nasr and Beba). Two concentrations of commercial deltamethrin 5% (200 and 400 ppm) were tested by AIT on R. (B.) annulatus. The mortality of ticks at 400 ppm was higher than 200 ppm which was not reached to 100%. Ehnasia tick population was the least susceptible to deltamethrin while Alfashin population was the most susceptible one. Different deltamethrin concentrations (0.000625, 0.00125, 0.0025, 0.005, 0.010 and 0.020) were used in LPT to evaluate the larval mortality. The highest LC50 appeared in Ehanasia tick population (0.01) with 95% confidence interval (0.005 to 0.01) meanwhile, the lowest LC50 appeared in Alfashin (0.0006) with 95%CI (0.0005 to 0.0007). Thus, it is possible to deduce that one population was considered susceptible; seven populations were resistant level I and three populations were resistant level II. Additionally, PCR revealed that the resistant alleles (R) appeared in two localities (Ehnasia and Aldiabia). It was concluded that at least two tick populations of the investigated locations were of possible resistant to deltamethrin.

Immunoprotective Effect of Chitosan Particles on Hymenolepis nana - Infected Mice.

Hymenolepis nana is the most commonly known intestinal cestode infecting mainly human. This study aimed to investigate the potential effect of chitosan particles (CSP) to enhance the immune system against H. nana infection. Determination of worm burden, egg output, histopathological changes, oxidative stress markers (lipid peroxidation and reduced glutathione), goblet (GCs) and mucosal mast cells (MMCs) counts in intestinal ileum was performed. In addition, levels of intestinal mRNA expression of interleukin (IL)-4, IL-9, stem cell factor (SCF), type I and II interferons (IFN)-α/ γ, tumour necrosis factor (TNF)-α, mucin 2 (MUC2) and inducible nitric oxide synthase (iNOs) were investigated using real-time PCR. The results indicated induced reductions in adult worm and egg counts in infected mice after CSP treatment. This was associated with improvement in tissue morphometric measurements and oxidative stress which were altered after infection. Expression levels of iNOs, IFN-α, IFN-γ, TNF-α and IL-9 were decreased by CSP. Conversely, expression levels of MUC2, IL-4 and SCF increased compared to infected untreated group. In addition, GCs and MMCs counts were normalized by CSP. In conclusion, this study could indicate the immunoprotective effect of CSP against H. nana infection. This was characterized with Th2 anti-inflammatory responses.

In vitro and in vivo effect of Citrus limon essential oil against sarcoptic mange in rabbits.

The effect of lemon oil (Citrus limon) on Sarcoptes scabiei var. cuniculi was evaluated in vitro and in vivo. The mite samples were collected from naturally infected rabbits. The lemon oil was prepared in six concentrations by dilution with distilled water (2.5, 5, 10, 20, 50, and 100 %). In vitro application was done in five replicates for each concentration in petri dishes in the laboratory. The treated mites were observed at 1, 12, and 24 h post application (PA) for lemon oil effect. In addition, oxidative stress profile was evaluated for the treated mite. Dependent on in vitro results, 20 % lemon oil was used in vivo trial. Twenty-four naturally infected rabbits were divided into three groups of eight: 20 % lemon oil, deltamethrin, and untreated control. The infected parts of rabbits were treated topically once a week for four successive weeks. In vitro application results showed that lemon oil 10 and 20 % diluted in water caused mortality to 100 % of mites after 24 h PA. The oxidative stress profile revealed that mites treated with 20 % lemon oil had significantly (P < 0.05) higher hydrogen peroxide and malondialdehyde when compared with mites treated with deltamethrin or distilled water. In vivo application of 20 % lemon oil on naturally infected rabbits showed complete recovery from clinical signs, absence of mite in microscopic examination from the second week of treatment. In addition, productive performance was significantly better than infected untreated group. Also, the treated tissue showed stoppage of scale formation and hair growth faster than deltamethrin-treated rabbits. Consequently, lemon oil has remarkable miticidal activity in vitro and in vivo applications.

Epidemiology and public health significance of Cryptosporidium isolated from cattle, buffaloes, and humans in Egypt.

The epidemiology and public health significance of Cryptosporidium species and genotypes were investigated in Beni-Suef Governorate, Egypt. A total of 610 animal fecal samples (480 from cattle and 130 from buffaloes) beside 290 stool samples from humans were collected in the period between January and December 2014. Based on the microscopic examination, the overall estimated prevalence of Cryptosporidium spp. in cattle, buffaloes, and humans was 10.2, 12.3, and 19 %, respectively. The highest detection rates were in calves less than 2 months of age (17.1 %) and diarrheic animals (13.0 %). Likewise in humans, the highest prevalence of Cryptosporidium was in infants (31.3 %) and diarrheic individuals (21.1 %). The gender distribution in humans denoted that Cryptosporidium was reported more frequently in males (21.7 %) than females (14.5 %). Based on the molecular characterization of Cryptosporidium, Cryptosporidium oocyst wall protein (COWP) and gp60 genes were successfully amplified in 36 out of 50 samples subjected to genotyping. Restriction fragment length polymorphism (RFLP) analysis of the COWP fragments revealed that Cryptosporidium parvum was the only species detected in cattle (12 isolates) and buffaloes (4 isolates), while in humans, the detected species were Cryptosporidium hominis (15 isolates) and C. parvum (5 isolates). Sequence analysis of the gp60 gene identified the subtype IIdA20G1 within C. parvum isolated from both animals and humans. The common occurrence of zoonotic subtypes of C. parvum in cattle and buffaloes highlights the potential role of these animals as significant reservoirs of infection to humans. Also, the presence of C. hominis and C. parvum in humans indicates that both anthroponotic and zoonotic pathways are expected.

Effect of Allium sativum and Allium cepa oils on different stages of Boophilus annulatus.

The study is aimed to investigate the acaricidal effect of Allium sativum (garlic) and Allium cepa (onion) oils on different stages of Boophilus annulatus hard tick. Engorged B. annulatus females were collected from naturally infected cattle. A number of engorged ticks were incubated at 28 °C and 85 % relative humidity to lay eggs, which were incubated to obtain larvae that were used in the study. The used garlic and onion oils were prepared by steam distillation and were analyzed by gas chromatography. These oils were dissolved in ethanol, methanol alcohols, and, partially, in water. The oils were tested in different concentrations; 1, 2, 5, 10, and 20 %. These concentrations were applied on adult ticks by adult immersion test; on larvae by larval immersion technique and on eggs. The 20, 10, and 5 % of garlic oil dilutions in ethanol and methanol killed all adult ticks and larvae within 24 h. Similar results were obtained for 10 and 20 % garlic oil dissolved in water. The effect of 10 % aqueous solution of garlic oil on embryonated eggs was clear as its addition to these eggs led to their in ability to hatch, deformity in shape, and change in color. The 10 and 20 % onion oil in ethanol and methanol alcohols killed 76-86 % of the adult ticks within 72 h post-application. While, all larvae died within 24 h postsubjected to these two concentrations. These concentrations (10 and 20 %) of onion oil in water killed 56-80 % of the treated ticks. Moreover, 10 % aqueous solution of onion oil prevented hatching of embyonated eggs. We concluded that garlic and onion oils have acaricidal effect on all stages of B. annulatus at concentrations higher than 5 %. Only garlic oil could kill 100 % of adult ticks at concentrations from 5 % in alcohols.