Emerging alternatives to traditional anthelmintics: the in vitro antiparasitic activity of silver and selenium nanoparticles, and pomegranate (Punica granatum) peel extract against Haemonchus contortus.

Haemonchus contortus (H. contortus) is one of the most prevalent gastrointestinal nematodes, causing health problems and economic losses in ruminants. Nanotechnology holds great promise as a field of science, with potential applications in veterinary medicine. This study investigated the in vitro anthelmintic activity of silver nanoparticles (AgNPs), selenium nanoparticles (SeNPs), and pomegranate peel extract (Punica granatum; PPE) on different stages of H. contortus: eggs, larvae, and adults. The in vitro anthelmintic efficacy was evaluated using the egg hatching inhibition assay (EHA), the third larval stage paralysis assay (LPA), and the adult worm motility inhibition assay (WMI). Six dilutions of PPE were utilized for EHA, LPA, and WMI, ranging from 0.25 to 6 mg/ml. AgNPs dilutions ranged from 0.00001 to 1.0 µg/ml for EHA and LPA and 1 to 25 µg/ml for WMI. SeNPs were utilized at dilutions of 1, 5, 10, and 15 µg/ml for EHA, LPA, and WMI. The results showed that the lowest concentration of AgNPs, SeNPs, and PPE significantly inhibited egg hatching. To further assess larvicidal activity, AgNPs at the highest concentration of 1 µg/ml induced a strong larvicidal effect, as did SeNPs at the lowest concentration. On the contrary, PPE displayed a significant larvicidal effect at 1 mg/ml compared to the control. The percentage mortality of adult H. contortus was measured as follows (mortality (%) = the number of dead adult H. contortus/total number of adult H. contortus per test × 100). The death of the adult H. contortus was determined by the absence of motility. Adult H. contortus mortality percentage was also significantly affected by all three agents when compared to the control. The AgNPs, SeNPs, and PPE have effective antiparasitic activity on gastrointestinal parasitic nematodes. These results provide evidence of the excellent antiparasitic properties of AgNPs, SeNPs, and PPE, demonstrating their effectiveness in controlling eggs, larvae, and adult H. contortus in vitro.

Anthelmintic activity of pomegranate peel extract (Punica granatum) and synthetic anthelmintics against gastrointestinal nematodes in cattle, sheep, goats, and buffalos: in vivo study.

Parasitic gastroenteritis (PGE) is one of the most important parasitic diseases that causes economic losses and health problems in ruminants. PGE causes a drop in milk, meat, and wool production in addition to decreasing animal fertility and sometimes leading to animal death. Conventional anthelmintics used for animal treatment are expensive, especially for farmers in developing countries. Moreover, the concern of anthelmintic resistance to these synthetic drugs is rising. Therefore, this study aimed to evaluate the anthelmintic activity of plant extract pomegranate (Punica granatum L) peel extract (PPE) against PGE infestations among ruminants. A total of 120 ruminants of different species (20 cattle, 12 buffalos, 68 sheep, and 20 goats) were examined for PGE eggs in their fecal samples. The animals under experiment were divided into four groups: the first group (negative control) was not given any drugs, the second group was given ivermectin (0.5 ml/25 kg bwt) (positive control 1), the third group was given albendazole (2.5 mg active principle/kg bwt) (positive control 2), and the fourth group was given PPE (200 mg/kg bwt). Fecal egg count (FEC) was performed on day 0 prior to the 1st dose of treatment. On day 15, an additional treatment (with the same doses) was administered and FEC was performed on days 7 and 21. Our results showed that on the 7th day of the experiment, there was an increase in FEC in the negative control group by 5%, while in the second, third, and fourth groups, there was a decrease in FEC with 95%, 90%, and 85% respectively. On the 21st day (7 days from the second dose), there was an increase in FEC in the control group by a 10% and 100% reduction in FEC in both the second and third groups. While in the fourth group, there was a decrease in FEC by 97%. In conclusion, PPE could be used as a safe, cheap, and effective natural anthelmintic against PGE.

The Role of Silver Nanoparticles in a Treatment Approach for Multidrug-Resistant Salmonella Species Isolates.

PURPOSE: The main objective of this study is to investigate the antibacterial activity of silver nanoparticles (AgNPs) against multidrug-resistant Salmonella isolates recovered from diarrheic  sheep and goats. METHODS: This study used chemical reduction synthesis of AgNPs to evaluate their antimicrobial effects by estimation of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for each isolate using the microplate dilution method and tetrazolium salt reduction test to detect the viability percentage. In vivo treatment efficacy was assessed in mice by determining the viable count of Salmonella Enteritidis recovered from feces and by hematologic, biochemical and histopathologic examinations to confirm that use of AgNPs has no toxic or pathologic effects and to evaluate its ability in tissue regeneration following treatment. RESULTS: All recovered strains were identified as MDR with a prevalence of 4% and 3.6% in sheep and goats, respectively. The results of TEM, DLS, Zeta potential, and FTIR revealed typical characteristics of the synthesized AgNPs. Silver nanoparticles showed antibacterial activity against all recovered strains with MIC of ≤0.02-0.313 µg/mL (mean average 0.085±0.126 µg/mL) and MBC of 0.078-1.250 µg/mL (average 0.508±0.315 µg/mL). In vivo efficacy of AgNPs was observed by a reduction in the number of viable S. Enteritidis recovered from feces in an S. Enteritidis infected mouse model, with complete shedding stopping between treatment days 4 and 6. Hematologic, serum biochemical, and histopathologic analyses proved the ability of AgNPs to suppress inflammatory reaction caused by S. Enteritidis infection. CONCLUSION: The study proved the effective ability of AgNPs to fight MDR Salmonella spp. in vitro and in vivo without adverse effects.

Isolation and molecular characterization of Mycoplasma spp. in sheep and goats in Egypt.

BACKGROUND AND AIM: Different species of Mycoplasma are associated with many pathological problems in small ruminants including respiratory manifestation, this problem results in significant losses, especially in African countries. This study aimed to (I) study some epidemiological aspects of Mycoplasma species infections in Egyptian sheep and goats at Giza Governorate, (II) diagnosis of Mycoplasma species affections using bacterial isolation and identification, (III) apply the polymerase chain reaction (PCR) for typing of different Mycoplasma species, and (IV) illustrate the phylogenetic tree for the isolated Mycoplasma species and other species from GenBank using the purified PCR product. MATERIALS AND METHODS: A total of 335 samples were collected from sheep and goats from Giza Governorate in Egypt as 142 nasal swabs from clinically affected animals, 167 pneumonic lungs, 18 samples from tracheal bifurcation, and 8 samples by bronchial wash were cultured on pleuropneumonia-like organisms (PPLOs) media for cultivation of Mycoplasma species. PCR and sequencing and phylogenetic analysis were adopted to identify and classify the isolated Mycoplasma species. RESULTS: A total of 24 Mycoplasma isolates were isolated on PPLO media, identified by biochemical tests, and confirmed and typed by PCR using specific primers. 10 isolates were confirmed as Mycoplasma arginini, four isolates as Mycoplasma ovipneumoniae by PCR, and 10 isolates as undifferentiated Mycoplasma species. A purified isolate of M. arginini and M. ovipneumoniae was sequenced and phylogenetic analysis was illustrated. CONCLUSION: M. arginini and M. ovipneumoniae are prevalent in Egyptian sheep and goats. Further studies on M. arginini are required due to its high frequency of isolation from pneumonic sheep and goats and also from animals suffer from different respiratory manifestations.

Morphological and molecular identification of the brown dog tick Rhipicephalus sanguineus and the camel tick Hyalomma dromedarii (Acari: Ixodidae) vectors of Rickettsioses in Egypt.

AIM: Rickettsioses have an epidemiological importance that includes pathogens, vectors, and hosts. The dog tick Rhipicephalus sanguineus and the camel tick Hyalomma dromedarii play important roles as vectors and reservoirs of Rickettsiae. The aim of this study was to determine the prevalence of Rickettsiae in ixodid ticks species infesting dogs and camels in Egypt, in addition to, the morphological and molecular identification of R. sanguineus and H. dromedarii. MATERIALS AND METHODS: A total of 601 and 104 of ticks' specimens were collected from dogs and camels, respectively, in Cairo, Giza and Sinai provinces. Hemolymph staining technique and OmpA and gltA genes amplification were performed to estimate the prevalence rate of Rickettsiae in ticks. For morphological identification of tick species, light microscope (LM) and scanning electron microscope (SEM) were used. In addition to the phylogenetic analyses of 18S rDNA, Second internal transcript spacer, 12S rDNA, cytochrome c oxidase subunit-1, and 16S rDNA were performed for molecular identification of two tick species. RESULTS: The prevalence rate of Rickettsiae in ticks was 11.6% using hemolymph staining technique and 6.17% by OmpA and gltA genes amplification. Morphological identification revealed that 100% of dogs were infested by R. sanguineus while 91.9% of camels had been infested by H. dromedarii. The phylogenetic analyses of five DNA markers confirmed morphological identification by LM and SEM. The two tick species sequences analyses proved 96-100% sequences identities when compared with the reference data in Genbank records. CONCLUSION: The present studies confirm the suitability of mitochondrial DNA markers for reliable identification of ticks at both intra- and inter-species level over the nuclear ones. In addition to, the detection of Rickettsiae in both ticks' species and establishment of the phylogenetic status of R. sanguineus and H. dromedarii would be useful in understanding the epidemiology of ticks and tick borne rickettsioses in Egypt.