Mohamed Ibrahim Shaalan

BACKGROUND: members of the genus Sarcocystis are intracellular obligate protozoan parasites classified within the phylum Apicomplexa and have an obligate heteroxenous life cycle involving two hosts. A more comprehensive understanding of the prevalence and geographic range of different Sarcocystis species in marine ecosystems is needed globally and nationally. Hence, the objective of this study was to document the incidence of Sarcocystis infection in sharks within the aquarium ecosystem of Egypt and to identify the species through the characterization of the SSU rDNA gene.

METHODS: All organs of the mako shark specimen underwent macroscopic screening to detect the existence of a Sarcocystis cyst. Ten cysts were collected from the intestine and processed separately to extract the genomic DNA. The polymerase chain reaction (PCR) was accomplished by amplifying a specific 18S ribosomal RNA (rRNA) gene fragment. Subsequently, the resulting amplicons were subjected to purification and sequencing processes.

RESULTS: Macroscopic examination of the mako shark intestinal wall sample revealed the presence of Sarcocystis cysts of various sizes and shapes, and sequencing of the amplicons from Sarcocystis DNA revealed a 100% nucleotide identity with the sequence of Sarcocystis tenella recorded from sheep in Iran; The mako shark sequence has been deposited in the GeneBank with the accession number OQ721979. This study presents the first scientific evidence demonstrating the presence of the Sarcocystis parasite in sharks, thereby documenting this specific marine species as a novel intermediate host in the Sarcocystis life cycle.

CONCLUSIONS: This is the first identification of Sarcocystis infection in sharks, and we anticipate it will be an essential study for future screenings and establishing effective management measures for this disease in aquatic ecosystems.

BACKGROUND: Pseudomonas putida is a pathogenic bacterium that induces great losses in fishes, including Nile tilapia (Oreochromis niloticus). Currently, the application of nanomaterials in aquaculture practices has gained more success as it endows promising results in therapies compared to traditional protocols.

OBJECTIVE: Therefore, the current perspective is considered the first report to assess the anti-bacterial efficacy of titanium dioxide nanogel (TDNG) against Pseudomonas putida (P. putida) in Nile tilapia.

METHODS: The fish (n = 200; average body weight: 47.50±1.32 g) were allocated into four random groups (control, TDNG, P. putida, and TDNG + P. putida), where 0.9 mg/L of TDNG was applied as bath treatment for ten days.

RESULTS: Outcomes revealed that P. putida infection caused ethological alterations (surfacing, abnormal movement, and aggression) and depression of immune-antioxidant variables (complement 3, lysozyme activity, total antioxidant capacity, superoxide dismutase, and reduced glutathione content). Additionally, a substantial elevation in hepatorenal biomarkers (aspartate and alanine aminotransferases and creatinine) with clear histopathological changes and immuno-histochemical alterations (very weak BCL-2 and potent caspase-3 immuno-expressions) were seen. Surprisingly, treating P. putida-infected fish with TDNG improved these variables and obvious restoration of the tissue architectures.

CONCLUSION: Overall, this report encompasses the key role of TDNG as an anti-bacterial agent for controlling P. putida infection and improving the health status of Nile tilapia.

Various kinds of pets have been known to contract the ectoparasite Sarcoptes scabiei. Current acaricides are becoming less effective because of the resistance developed by the mite besides their adverse effects on the general activity and reproductive performance of domestic pets. For this reason, the present study aims to discover a novel and safe approach using silver and gold nanoparticles to fight Sarcoptic mange in rabbits as well as to explain their mechanism of action. 15 pet rabbits with clinical signs of Sarcoptic mange that were confirmed by the microscopic examination were used in our study. All rabbits used in this study were assessed positive for the presence of different developing stages of S. scabiei. Three groups of rabbits (n = 5) were used as follows: group (1) didn't receive any treatment, and group (2 and 3) was treated with either AgNPs or GNPs, respectively. Both nanoparticles were applied daily on the affected skin areas via a dressing and injected subcutaneously once a week for 2 weeks at a dose of 0.5 mg/kg bwt. Our results revealed that all rabbits were severely infested and took a mean score = 3. The skin lesions in rabbits that didn't receive any treatments progressed extensively and took a mean score = of 4. On the other hand, all nanoparticle-treated groups displayed marked improvement in the skin lesion and took an average score of 0-1. All NPs treated groups showed remarkable improvement in the microscopic pictures along with mild iNOS, TNF-α, and Cox-2 expression. Both nanoparticles could downregulate the m-RNA levels of IL-6 and IFγ and upregulate IL-10 and TGF-1β genes to promote skin healing. Dressing rabbits with both NPs didn't affect either liver and kidney biomarkers or serum Ig levels indicating their safety. Our residual analysis detected AgNPs in the liver of rabbits but did not detect any residues of GNPs in such organs. We recommend using GNPs as an alternative acaricide to fight rabbit mange.

Pseudomonas aeruginosa (P. aeruginosa) is a serious zoonotic pathogen threaten the poultry industry causing severe economic losses therefor, this study aimed to isolation, phenotypic, molecular identification of P. aeruginosa from different avian sources (chickens, turkey, pigeons, table eggs, and dead in shell chicken embryos), from different Egyptian governorates (Giza, Qalubia, Beheira, El-Minya, and Al-Sharqia) with applying of antibiotic sensitivity test on all P. aeruginosa isolates. Highly resistant isolates (n = 49) were subjected to molecular identification of P. aeruginosa with detection of resistant genes including carbapenemase-encoding genes blaKPC, blaOXA-48, and blaNDM. On the base of molecular results, a highly resistant P. aeruginosa strain was tested for its pathogenicity on day old specific pathogen free (SPF) chicks. Also, in vitro experiment was adopted to evaluate the efficacy of silver nanoparticles (Ag-NPs) against highly antibiotic-resistant P. aeruginosa strains. The overall isolation percentage was from all examined samples were 36.2% (571/1,576) representing 45.2% (532/1,176) from different birds' tissues and 39/400 (9.7%) from total egg samples. Some of isolated strains showed multidrug resistance (MDR) against kanamycin, amoxicillin, amoxicillin-clavulanic acid, neomycin, chloramphenicol, vancomycin, cefotaxime clavulanic acid, lincomycin-spectinomycin, co-trimoxazole, cefoxitin, gentamycin, and doxycycline. These MDR strains were also molecularly positive for ESBL and carbapenemase-encoding genes. MDR strain showed high pathogenicity with histopathological alterations in different organs in challenged birds. Main histopathological lesions were necrosis of hepatocytes, renal tubular epithelium, and heart muscle bundles. The MDR strain showed in vitro sensitivity to Ag-NPs. In conclusion, MDR P. aeruginosa is a serious pathogen causing high morbidity, mortality, and pathological tissue alterations. Ag NPs revealed a promising in vitro antimicrobial sensitivity against MDR P. aeruginosa and further in vivo studies were recommended.

Pseudomonas aeruginosa (P. aeruginosa) is a serious zoonotic pathogen threaten the poultry industry causing severe economic losses therefor, this study aimed to isolation, phenotypic, molecular identification of P. aeruginosa from different avian sources (chickens, turkey, pigeons, table eggs, and dead in shell chicken embryos), from different Egyptian governorates (Giza, Qalubia, Beheira, El-Minya, and Al-Sharqia) with applying of antibiotic sensitivity test on all P. aeruginosa isolates. Highly resistant isolates (n = 49) were subjected to molecular identification of P. aeruginosa with detection of resistant genes including carbapenemase-encoding genes blaKPC, blaOXA-48, and blaNDM. On the base of molecular results, a highly resistant P. aeruginosa strain was tested for its pathogenicity on day old specific pathogen free (SPF) chicks. Also, in vitro experiment was adopted to evaluate the efficacy of silver nanoparticles (Ag-NPs) against highly antibiotic-resistant P. aeruginosa strains. The overall isolation percentage was from all examined samples were 36.2% (571/1,576) representing 45.2% (532/1,176) from different birds' tissues and 39/400 (9.7%) from total egg samples. Some of isolated strains showed multidrug resistance (MDR) against kanamycin, amoxicillin, amoxicillin-clavulanic acid, neomycin, chloramphenicol, vancomycin, cefotaxime clavulanic acid, lincomycin-spectinomycin, co-trimoxazole, cefoxitin, gentamycin, and doxycycline. These MDR strains were also molecularly positive for ESBL and carbapenemase-encoding genes. MDR strain showed high pathogenicity with histopathological alterations in different organs in challenged birds. Main histopathological lesions were necrosis of hepatocytes, renal tubular epithelium, and heart muscle bundles. The MDR strain showed in vitro sensitivity to Ag-NPs. In conclusion, MDR P. aeruginosa is a serious pathogen causing high morbidity, mortality, and pathological tissue alterations. Ag NPs revealed a promising in vitro antimicrobial sensitivity against MDR P. aeruginosa and further in vivo studies were recommended.

Avian infectious bronchitis is one of the most common viral infections in chickens affecting all ages. The tropism of infectious bronchitis virus (IBV) strains became broader and more variable posing major implications for the effective control of IBV infection. In this study, two IBV viruses representing classic and variant strains were inoculated intranasally into day-old SPF chicks (10 EID/0.2 ml/bird). Clinical signs were observed for 15 days post-infection (DPI). Five chicks from each group were euthanized at 2, 4, 6, 8, 10, 12, and 15 DPI for histopathology and virus antigen detection by IHC and quantitative rRT-PCR. Results revealed that both classic and variant IBV strains induced mild clinical signs with no mortalities and fewer various histopathological lesions in infected SPF chickens. Although the viruses were detected by rRT-PCR up to 12 DPI, the affected tissues showed regeneration after 10 DPI with IHC revealing no IBV antigen. In summary, no differences were found in the behaviour of both IBV isolates in chickens. The broad tissue tropism for both IBV strains as indicated by viral antigen detection in various organs with no clinical or gross lesion suggest that the main cause of death in IBV infection under field conditions occurs as a result of complication with secondary infections rather single IBV infection. Due to positive immunostaining in the bursa, it is thought that IBV infection has immunosuppressive consequences, hence further study is required to validate this impact.

Avian infectious bronchitis is one of the most common viral infections in chickens affecting all ages. The tropism of infectious bronchitis virus (IBV) strains became broader and more variable posing major implications for the effective control of IBV infection. In this study, two IBV viruses representing classic and variant strains were inoculated intranasally into day-old SPF chicks (10 EID/0.2 ml/bird). Clinical signs were observed for 15 days post-infection (DPI). Five chicks from each group were euthanized at 2, 4, 6, 8, 10, 12, and 15 DPI for histopathology and virus antigen detection by IHC and quantitative rRT-PCR. Results revealed that both classic and variant IBV strains induced mild clinical signs with no mortalities and fewer various histopathological lesions in infected SPF chickens. Although the viruses were detected by rRT-PCR up to 12 DPI, the affected tissues showed regeneration after 10 DPI with IHC revealing no IBV antigen. In summary, no differences were found in the behaviour of both IBV isolates in chickens. The broad tissue tropism for both IBV strains as indicated by viral antigen detection in various organs with no clinical or gross lesion suggest that the main cause of death in IBV infection under field conditions occurs as a result of complication with secondary infections rather single IBV infection. Due to positive immunostaining in the bursa, it is thought that IBV infection has immunosuppressive consequences, hence further study is required to validate this impact.

Cattle Egret (Bubulcus ibis) is one of the most well-known herons in Egypt. It is called the friend of the farmer because it benefits farmers and helps them get rid of insects and worms. It acts as a reservoir for many diseases. Few researchers have discussed the significance of parasitic diseases that affect this wild bird and may lead to mortalities among the population especially the importance of vital organs such as kidneys. Therefore, this study aimed to spotlight parasitic infection-affected herons in Egypt and consider the risks to this beneficial bird. During this study, 23 Bubulcus ibis were captured after their death from Abou Rewash Giza Governorate, Egypt, during the period from February to September (2022). Renicola species (spp.) and Apharyngostrigea spp. are two important digenean parasites that were recovered from the kidneys, and small intestine of the heron Cattle Egret (Bubulcus ibis) with an infection rate of (17.2%) and (11.8%) respectively. Histopathological techniques were used to assess tissue alterations while light microscopy and molecular assays were used to assess the parasites. The parasites' morphological and morphometrical characteristics, as well as polymerase chain reaction and sequencing assays (mitochondrial sections), were investigated for the first time in Egypt. These parasites were given in-depth illustrations and drawings. The distinctive qualities of the two species were discussed. As the first record from Egypt, the nucleotide sequences discovered in this work have been uploaded into the GenBank database (accession numbers: OR021986 and OQ955829). Microscopically, the renal blood vessels had vasculitis, necrosis, and other degenerative alterations. Further research analyzing the health of various heron spp. and environmental deterioration can help to close information gaps about the interactions between parasites, their hosts, and environmental health.

The efficacy of silver nanoparticles (AgNPs) was tested in vitro against three different fish viruses, causing significant economic damage in aquaculture. These viruses were the spring viraemia of carp virus (SVCV), European catfish virus (ECV), and Ictalurid herpesvirus 2 (IcHV-2). The safe concentration of AgNPs that did not cause cytotoxic effects in EPC cells proved to be 25 ng/mL. This dose of AgNPs decreased significantly (5-330×) the viral load of all three viruses in three different types of treatments (virus pre-treatment, cell pre-treatment, and cell post-treatment with the AgNPs). In a higher concentration, the AgNPs proved to be efficient against ECV and IcHV-2 even in a delayed post-cell-treatment experiment (AgNP treatment was applied 24 h after the virus inoculation). These first in vitro results against three devastating fish viruses are encouraging to continue the study of the applicability of AgNPs in aquaculture in the future.

The efficacy of silver nanoparticles (AgNPs) was tested in vitro against three different fish viruses, causing significant economic damage in aquaculture. These viruses were the spring viraemia of carp virus (SVCV), European catfish virus (ECV), and Ictalurid herpesvirus 2 (IcHV-2). The safe concentration of AgNPs that did not cause cytotoxic effects in EPC cells proved to be 25 ng/mL. This dose of AgNPs decreased significantly (5-330×) the viral load of all three viruses in three different types of treatments (virus pre-treatment, cell pre-treatment, and cell post-treatment with the AgNPs). In a higher concentration, the AgNPs proved to be efficient against ECV and IcHV-2 even in a delayed post-cell-treatment experiment (AgNP treatment was applied 24 h after the virus inoculation). These first in vitro results against three devastating fish viruses are encouraging to continue the study of the applicability of AgNPs in aquaculture in the future.

BACKGROUND: Asthma is a chronic inflammatory lung disease that universally affects millions of people. Despite numerous well-defined medications, asthma is poorly managed. This study aims to clarify the potential therapeutic effect of Dapagliflozin (DAPA) against lung inflammation, oxidative stress, and associated bronchospasm in OVA-sensitized rat asthma model.

RESEARCH DESIGN AND METHODS: Twenty-five rats were allocated into (Control, Asthma, DEXA, DAPA, and DAPA+DEXA). All treatments were administered orally once a day for two weeks. The BALF levels of IL-17, TNFα, IL-1β, and MCP-1 were determined to assess airway inflammation. For oxidative stress determination, BALF MDA levels and TAC were measured. The BALF S100A4 level and NO/sGC/cGMP pathway were detected. Lung histopathological findings and immunohistochemical investigation of eNOS and iNOS activities were recorded.

RESULTS: DAPA significantly reduced () airway inflammatory-oxidative markers (IL-17, TNFα, IL-1β, MCP1, and MDA), but increased () TAC, and mitigated bronchospasm by activating NO/sGC/cGMP and reducing S100A4 (). The biochemical and western blot studies were supported by histopathological and immunohistochemical investigations.

CONCLUSIONS: DAPA presents a new prospective possibility for future asthma therapy due to its anti-inflammatory, anti-oxidant, and bronchodilator properties. DAPA has the property of reducing Dexamethasone (DEXA)-associated unfavorable effects during asthma treatment.

Equid alphaherpesvirus 1 (EHV-1) is the most important virus causing pathological disorders in horses and causes persistent outbreaks of upper respiratory tract infection, ocular affections, abortion, and neurological disorders with high mortality in Arabian horses in Egypt. As EHV-1 is a very contagious disease, the quick and accurate diagnosis are important to broaden our understanding of EHV-1 in the field, implicate strong preventive, and control measures. Sixty-six samples collected over a period of 4 years were examined. Most of samples originated from Cairo and Giza governorates from respiratory, abortigenic and neurological outbreaks. EHV-1 was diagnosed in these clinical cases by immunohistochemistry using monoclonal antibody against EHV-1 glycoprotein B and molecular detection using gB, ORF33 specific real-time PCR. Molecular characterization of glycoprotein B (gB, ORF33) gene was applied for confirmation. Molecular characterization revealed that the ORF33 sequences of this study were identical. These sequences were closely related to the European EHV-1 strains. Furthermore, EHV-1 sequences in this study showed little to no differences for the amino acid sequences compared to previously published sequences. This study would be valuable for monitoring of EHV-1 infection in Egypt and determining the gB gene sequence of newly identified EHV-1 field strains which is the most conserved region in the viral DNA and frequently used as a target for diagnostic PCR protocols for the future outbreaks.

Infectious laryngotracheitis (ILT) is an economically crucial respiratory disease of poultry that affects the industry worldwide. Vaccination is the principal tool in the control of the disease outbreak. In an earlier study, we comprehensively characterized the circulating strains in Egypt and identified both CEO-like and recombinant strains are dominant. Herein, we investigated the pathogenicity of two virulent strains representing the CEO-like (Sharkia_2018) and recombinant strain (Qalubia_2018). Additionally, we evaluated the efficacy of different commercial vaccines (HVT-LT, CEO, and TCO) against the two isolates in terms of the histopathological lesion scores and the viral (gC) gene load. A total of 270 White Leghorn-specific pathogen-free male chicks were divided into nine groups of 30 birds, each housed in separate isolators. Birds were distributed as follows; one group was non-vaccinated, non-challenged, and served as a negative control. Two groups were non-vaccinated and infected with the two isolates of interest and served as a positive control to test the pathogenicity. Six groups were vaccinated and challenged; two groups were vaccinated with vector vaccine at one day old. The other four groups were vaccinated with either the CEO- or TCO- vaccine (two groups each) at four weeks of age. Three weeks after vaccination, birds were infected with the virulent ILTV isolates. The larynx, trachea, and harderian gland samples were taken at 1, 3, and 7 days post-infection for histopathological lesion score and molecular detection. Notably, The recombinant strain was more virulent and pathogenic than CEO-like ILTV strains. Moreover, the TCO vaccine was less immunogenic than the vector and CEO vaccines.

Silica nanoparticles (SiNPs) are among the non-toxic nanoparticles (NPs) that have magnetic capabilities. It is hypothesized that SiNPs may be able to reduce toxic effects exerted by a mixture of lead (Pb) and mercury (Hg) in African catfish Clarias gariepinus. The in vitro magnetic potential of SiNPs to absorb Pb and Hg was tested. Fish (N = 240) were divided into four groups in triplicates for 30 days. The first group served as control and the second group (SiNPs) was exposed to 1/10 of 96 h LC50 of SiNPs (14.45 mg/L). The third group (HMM) was exposed to 1/10 of 96-h LC50 of a mixture of mercury chloride (HgCl) and lead chloride (PbCl) equal to 0.04 mg/ L and 23.1 mg/L. The fourth group (SiNPs+ HMM) was exposed to a suspension composed of SiNPs, HgCl, and PbCl at the same concentrations as the third group. Results showed that fish exposed to heavy metals revealed the following consequences; a significant decrease in hematological, immunological (complement-3 and nitric oxide), and antioxidants (total antioxidant capacity, glutathione peroxidase, superoxide dismutase, and catalase) indices, down-regulation of IL-1β, IL-8, TGF-β, NF-κβ, HSP70, and Hepcidin genes, the highest mortality rate (48.33%), higher values of alkaline phosphatase, alanine, and aspartate aminotransferases, urea, creatinine, and branchial malondialdehyde, marked up-regulation of CC chemokine and CXC chemokines, and high HMs residues levels in muscles. Extensive pathology showed degeneration with diffuse vacuolation of hepatopancreatic cells and hemorrhage in the HMM group. Interestingly, the exposed group to SiNPs and HMM demonstrated a decline of HMs concentration in fish muscles and modulated the abovementioned parameters with the regeneration of histological alterations of liver and gills. Based on the study outcomes, we highlight the importance and the safety of SiNPs as a novel aqueous additive to alleviate HMs toxicity and recommend using SiNPs for enhancing fish performance for sustaining aquaculture without adverting safety of human health by their little accumulation in muscular tissue.

Neuroinflammation, a major component of many CNS disorders, has been suggested to be associated with diacetyl (DA) exposure. DA is commonly used as a food flavoring additive and condiment. Lately, silymarin (Sily) has shown protective and therapeutic effects on neuronal inflammation. The study aimed to explore the role of Sily in protecting and/or treating DA-induced neuroinflammation. Neuroinflammation was induced in rats by administering DA (25 mg/kg) orally. Results revealed that Sily (50 mg/kg) obviously maintained cognitive and behavioral functions, alleviated brain antioxidant status, and inhibited microglial activation. Sily enhanced IL-10, GDNF and Dyn levels, reduced IFN-γ, TNFα, and IL-1β levels, and down-regulated the MAPK pathway. Immunohistochemical investigation of EGFR and GFAP declared that Sily could conserve neurons from inflammatory damage. However, with continuing DA exposure during Sily treatment, oxidative stress and neuroinflammation were less mitigated. These findings point to a novel mechanism involving the Dyn/GDNF and MAPK pathway through which Sily might prevent and treat DA-induced neuroinflammation.

Marine bio-sourced chitosan nanoparticles (CSNP) are antimicrobial and immunomodulatory agents beneficial for fish medicine. Herein, dietary CSNP was investigated for the amelioration of the systemic inflammatory responses of an induced fish model. One hundred and forty-four rainbow trout were assigned to one pathogen-free and non-supplemented group (negative control), and three challenged groups: non-supplemented (positive control), CSNP-preventive, and CSNP-therapeutic. After a feeding experiment extended for 21 days, the organosomatic indices (OSI) and molecular aspects were assessed. After a challenge experiment extended for further 28 days, CSNP-therapeutic intervention was assessed on fish survival and systemic inflammatory responses on pathology, histo-morphology, and molecular aspects. With CSNP administration, OSI nonsignificantly decreased and the relative expression of targeted inflammatory-mediator genes was significantly increased. The CSNP-therapeutic fish showed an RPS of 80% as compared to the positive control group, and CSNP-therapeutic administration retained the highest gene expression augmentation up to 28 days after the challenge. Notably, the splenic reticulin fibers framework of the CSNP-therapeutic group retained the highest integrity among the groups during the infection. After recovery, reticulin fibers density in the CSNP-therapeutic samples was significantly higher than in the negative control group, which indicates high innate immunity. Thus, CSNP showed promising biotherapeutic features enhancing fish resistance against infections.

Leishmaniasis is a neglected zoonotic disease that poses significant veterinary and public health risks in developing countries. Dogs act as a reservoir host for leishmaniasis transmitted to humans. A total of 108 human cases of cutaneous leishmaniasis (CL) were identified in the Al-Houd Al-Marsoud Hospital in Cairo, Egypt, during 2018. Blood samples and skin biopsies were collected for further examination. Blood samples from 96 asymptomatic dogs were collected. All samples were subjected to molecular and phylogenetic analysis. Quantitative RT-PCR was used to measure the expression of genes related to mTOR signalling and inflammation in blood and tissue samples. The distribution pattern of human cases pointed to an endemic focus in North Sinai (66.67%). The prevalence of asymptomatic canine leishmaniasis was 66.60%. Histopathological examination of human skin lesions revealed a severe granulomatous inflammatory reaction, necrosis and ulceration. Moreover, leishmanial amastigotes could be detected in human tissue samples. Phylogenetic analysis revealed 100% identity of human isolates to Leishmania tropica (MN453682), and dog isolates to Leishmania infantum (MN453673), with 94.9% similarity between the two isolates. Gene expression related to mTOR signalling and inflammation in both species' samples confirmed a significant alteration of EIF4EBP1, CCR4 and INF-γ expression compared with control groups. In Egypt, increased incidence of asymptomatic carrier dogs acting as a significant reservoir host for Leishmania poses a public health hazard. Findings warrant further epidemiological investigation of CL in Egypt, as well as additional study of parasite differentiation and gene regulation.

Flavobacterial infections are among the causes of fish losses in farms with the emergence of antibiotic-resistant isolates. Silver nanoparticles (AgNPs) are known for their potent antimicrobial activity against different types of bacteria. In this study, we evaluated the antibacterial properties of AgNPs (diameter: 23 nm) against Flavobacterium johnsoniae infection in common carp Cyprinus carpio. The assays included both in vitro and in vivo antibacterial tests in addition to evaluation of cell toxicity effects on the fish cell lines. The in vitro results revealed potent inhibitory effects of AgNPs on the growth of F. johnsoniae with a minimum inhibitory concentration of 34 µg ml-1. Fish cell (epithelioma papulosum cyprini and koi carp fin) viability was 95-100% after exposure to 500 ng ml-1 (and lower concentrations) of AgNPs. In the exposure experiment, mortality rates decreased from 45% in the infected non-treated group to 30 and 15% in the intraperitoneal injection and immersion-treated groups, respectively. Neither of the treated groups showed any clinical signs or histopathological lesions. The single-dose treatment with AgNPs during early infection with F. johnsoniae aided in minimizing fish losses.

Purpose: The main objective of this study is to investigate the antibacterial activity of silver nanoparticles (AgNPs) against multidrug-resistant 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 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 . Enteritidis recovered from feces in an . 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 . Enteritidis infection.

Conclusion: The study proved the effective ability of AgNPs to fight MDR spp. in vitro and in vivo without adverse effects.

The rise of bacterial resistance to antibiotics is one of the great challenges of our age. One of the strategies to limit the development of antibiotics resistance is the investigation of alternative antimicrobials. As silver nanoparticles demonstrated a potent bactericidal activity in vitro, the aim of this study was to evaluate the in vivo antibacterial activity of silver nanoparticles against Aeromonas salmonicida subsp. salmonicida. Rainbow trout (n = 120) were divided into four groups of 30 fish each. First group was challenged with A. salmonicida (Positive control), the second group was challenged with A. salmonicida and exposed to silver nanoparticles by immersion for three hours (100 μg/L), the third group was challenged with A. salmonicida and intraperitoneally injected with silver nanoparticles (17 μg/mL) and the fourth group was sham-treated and served as a negative control group. At the 7th day post challenge, histopathology of the positive control group revealed the presence of bacterial aggregates in tissues with degenerative and necrotic changes, while at the 35th day post challenge, only liver necrosis persisted. Silver nanoparticles-treated and negative control groups did not show any clinical signs, mortalities or histopathological alterations and they were tested negative for A. salmonicida. The immersion in silver nanoparticles did not result in detectable residues of silver in the muscles 35 days after treatment. These findings demonstrate the antibacterial properties of silver nanoparticles against A. salmonicida infection. Therefore, they could be used for development of antibacterial agents in aquaculture.

Accurate and rapid identification of bacterial pathogens of fish is essential for the effective treatment and speedy control of infections. Massive mortalities in market-sized red tilapia (Oreochromis spp.) were noticed in mariculture concrete ponds in northern Egypt. Histopathological examination revealed marked congestion in the central vein of the liver with the presence of bacterial aggregates inside the lumen and in the vicinity of the central vein. A total of 12 isolates of streptococci were obtained from the moribund fish. This study documented the ability of the MicroSeq 500 16S bacterial sequencing method to accurately identify Streptococcus agalactiae and S. dysgalactiae mixed infections from moribund red tilapia that were difficult to be recognised by the commercial biochemical systems. The continuously decreasing cost of the sequencing technique should encourage its application in routine diagnostic procedures.