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Newcastle disease virus (NDV), a major pathogen of poultry worldwide, causes significant economic losses in the poultry industry. To characterize the ability of recently isolated virulent strains of NDV genotypes VI and VII to cause disease in quails, and to evaluate the efficacy of two NDV vaccines against such strains, Japanese quails were experimentally inoculated with either NDV genotype VI (Pigeon F-VI strain) or VII 1.1 (GHB-328 strain) with or without vaccination with inactivated NDV vaccine of genotype II (La Sota strain) or VII (KBNP strain). Mild to severe neurological signs developed in quails inoculated with the Pigeon F-VI strain from 3 to 14 days post infection (PI) and from 4 to 10 days PI in birds infected with the GHB-328 strain. The mortality rates were 46% and 33% for birds inoculated with NDV VI and NDV VII 1.1, respectively. The severity of histopathological changes depended on the viral isolates used. Vaccination with the La Sota or KBNP vaccine strain successfully protected quails against NDV-induced mortality and decreased the severity of clinical signs, pathological changes and cloacal viral shedding. This study showed that these virulent NDV isolates had mild to moderate pathogenicity in quails and that both vaccines protected against challenge with both virus strains. NDV vaccine genotype VII improved the level of protection against challenge with the VII 1.1 genotype compared with the classic vaccine, but failed to protect quails against challenge with the VI genotype.

The present study was intended for investigating the effect of lead acetate toxicity (5 or 10 mg lead acetate/liter water), Dunaliella algae (11 g/kg diet)-supplemented diet and their interaction i...

The present trial was performed to evaluate the effect of dietary incorporation of dried Spirulina, Chlorella and their mixture on the immune status of Nile tilapia (Oreochromis niloticus) before and after artificial infection with pathogenic bacterium Aeromonas hydrophila (A. hydrophila). Two hundred and forty fish were divided into four groups: (a) a control group fed on a basal diet only, (b) a second group fed on a diet containing Spirulina 15%, (c) a third group fed on a diet containing Chlorella 15%, and (d) a fourth group fed on a diet containing a mixture of both Spirulina 15% and Chlorella 15%. At the end of the experiment (9 weeks), the four groups were experimentally infected with A. hydrophila for 7 days. Antioxidant enzymes, lysozyme and bactericidal activities and histopathological changes were determined just before the challenge test and 7 days post‐challenge. Significant (p ≤ 0.05) increases in fish body protein% before the challenge test and increases in serum antioxidant enzymes, lysozyme and bactericidal activity in the Chlorella and algal mixture groups before and after the challenge test were observed. Spirulina, Chlorella and their mixture groups significantly decreased serum malondialdehyde compared to the control group before and after the challenge test. Using Spirulina, Chlorella and their mixture mitigated the necrotic and degenerative changes induced by A. hydrophila and revealed well‐developed and multiple melanomacrophage centres. Thus, dietary Spirulina, Chlorella and their mixture inclusion in Nile tilapia fish proved to have a protective effect against A. hydrophila infection.

The remediation of wastewater (WW) is a promising solution for limited water sources. This study aimed to evaluate rice straw (RS) and zeolite (Z) as bioadsorbents for the removal of pollutants, including heavy metals (HMs) (cadmium [Cd], nickel [Ni], and lead [Pb]) and malathion (PC), from WW and to assess the suitability of reusing remediated WW in fish rearing units. A total of 11 treatment groups with 3 replicates each were designed with different combinations of RS and/or Z for the treatment of real WW contaminated with HMs and malathion, where the WW remained in contact with the adsorbents for 24 h. Different remediated WWs were used for rearing Nile tilapia (Oreochromis niloticus), which were randomly allocated into 33 glass aquaria representing 11 treatments with 3 replicates each for 30 days. The best remediation efficiency was achieved using a mixture of whole RS (WRS), chopped RS (CRS), and Z (HM-PC-WRS-CRS-Z group), with removal percentages of 92%, 95%, 96%, and 99% for Cd, Ni, Pb, and malathion, respectively. The health status of the aquatic ecosystems was assessed through blood tests to characterize biochemical parameters and through pathological changes of cultured O. niloticus reared in treated WW. A significant (P ˂ 0.05) effect on the blood biochemistry of fish reared in treated WW was found and better biochemical and histologic architecture was observed than that of fish reared in untreated WW. A novel mixture of WRS, CRS, and Z could possibly be a promising low-cost adsorbent for wastewater treatment. Graphical abstract.

The present study aimed to investigate the toxic effect of aflatoxin B1 (AFB1) and the effectiveness of a biological mycotoxin binder (Nutritox®) in detoxifying aflatoxicosis. Ninety one-day-old chicks were chosen and divided into 3 groups. The first group received standard basal diet only, the second group received a basal diet with AFB1 (0.25 mg/kg diet), and the third group received a basal diet with AFB1 (0.25 mg/kg diet) and Nutritox® (1 kg/ton diet). AFB1 feed contamination significantly reduced growth performance and deteriorated FCR. Moreover, it significantly increased serum AST, ALT, and malondialdehyde and significantly decreased serum total protein, albumin, globulin, SOD, CAT activities and glutathione peroxidase gene expression as well. Aflatoxin residues were detected in the liver tissues. Furthermore, the liver and kidney of AFB1 treated group showed pathological changes. The supplementation of Nutritox® significantly reduced aflatoxin levels in the liver and counteracted the negative effects of AFB1.

Wound healing is a public health concern. Licorice gained a great attention for its antioxidant and anti-inflammatory properties which expand its valuable effects as a herbal medicine. In this study, we pointed out to the wound healing potential and the mechanism by which licorice alcoholic extract can modulate cutaneous wound healing through immune, antioxidant, histopathological, immunohistochemical (IHC) and molecular studies. 24 Wister rats were assigned into 3 groups (n = 8 each); control group, topical and oral supplied groups. Licorice extract administration significantly increased total and differential leucocyte counts, phagocytic activity of neutrophils, antioxidant biomarkers as superoxide dismutase (SOD), glutathione peroxidase activities (GPx) and reduced glutathione (GSH) content with a notable reduction in oxidative stress marker malondialdehyde (MDA). Moreover, histopathological findings detected complete re-epithelialization with increasing collagen synthesis while IHC results revealed a significant enhancement in the expression of α-SMA, PDGFR-α, FGFR1 and Cytokeratin 14 in licorice treated groups compared with the control group. Licorice extract supplementation accelerated wound healing by increasing angiogenesis and collagen deposition through up-regulation of bFGF, VEGF and TGF-β gene expression levels compared with the control group. UPLC-PDA-MS/MS aided to authenticate the studied Glycyrrihza species and recognized 101 potential constituents that may be responsible for licorice-exhibited potentials. Based on our observations we concluded that licorice enhanced cutaneous wound healing via its free radical-scavenging potential, potent antioxidant activities, and anti-inflammatory actions. Therefore, licorice could be used as a potential alternative therapy for wound injury which could overcome the associated limitations of modern therapeutic products.

We studied the impact of using fodder enriched with the cyanobacterium Spirulina platensis or the green alga Chlorella vulgaris or a consortium containing both of them on growth, biochemistry profile and immune status of the Nile tilapia (Oreochromis niloticus) fish. A set of 240 males challenged with Aeromonas hydrophila one week before the end of 9 weeks experimental period. The fodder supplemented with 15% (w/w) of Spirulina platensis, 15% of Chlorella vulgaris or 15% of their (1/1) mixture. Growth performance was measured all over 9 weeks. Challenge test was performed by infection with the pathogen after 8 weeks of growth. The serum total contents of protein, albumin, globulin, urea, creatinine and activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALK) and lactate dehydrogenase (LDH) were measured at 7 days before and after the infection. The results revealed significant increases in body weight of the Chlorella and their mixture (Chlorella and Spirulina) treated groups compared with the untreated control and the only Spirulina treated ones (p ≤ 0.05). Without infection, the serum total protein and serum globulin significantly increased by supplementation with the Spirulina, Chlorella and both compared to the uninfected control group. The serum albumin significantly decreased with Chlorella and the mixture supplementations. Lactate dehydrogenase (LDH) in the fish serum generally decreased with application of the enriched feed compared with the control. After infection, serum total protein and globulin significantly increased by the feed supplementations compared with the control group while the serum albumin insignificantly increased by supplementation with Chlorella and Spirulina. ALT, AST, ALK, LDH, urea and creatinine in the fish serum decreased with the microalgae additives compared with the control.

The infectivity of equine herpesvirus (EHV)-9 has been studied in different animal models including immunocompromised animals. The current study focused on the infectivity of EHV-9 in different mouse strains (C3H, C57BL, DBA, BALB/c-nu/nu, BALB/c and ICR) by intranasal inoculation of 2 × 106 plaque forming units (PFU). Various organs, including head and lungs, were collected 7 days post infection (dpi) to investigate microscopical lesions and the distribution of EHV-9 antigen. Immunopositivity of tissue sections was scored using ImageJ software. Open reading frame (ORF) 30 expression in lung tissues was quantified using quantitative reverse transcriptase polymerase chain reaction. Pathological examination revealed different degrees of rhinitis in the different mouse strains. Severe rhinitis was detected in C3H and BALB/c-nu/nu strains, moderate rhinitis was observed in C57BL and DBA strains and no lesions were detected in BALB/c mice. Immunopositivity for EHV-9 antigens was detected in the olfactory epithelium of C3H and BALB/c-nu/nu strains. Compared with C57BL, DBA, BALB/c-nu/nu, ICR and BALB/c strains, the C3H strain showed greater expression of EHV-9 antigens in the brain. The proportion of areas with high positive to positive immunoreactivity for EHV-9 were 7.57, 3.42, 3.12, 2.51, 1.79 and 0.03% for C3H, C57BL, DBA, BALB/c-nu/nu, ICR and BALB/c strains, respectively. The proportions of areas with low positive to negative immunoreactivity were 92.42, 96.70, 96.87, 97.48, 98.16 and 99.96%, respectively. The highest relative expression levels for EHV-9 ORF30 in the lungs were in C3H mice. No significant differences in the expression of ORF30 were observed in other strains. In conclusion, of the strains examined, C3H, C57BL, DBA, BALB/c-nu/nu and ICR were the most susceptible to EHV-9 infection, and the BALB/c strain was less susceptible.

Encephalitis in hamsters, which was induced by equine herpesvirus (EHV)–9, EHV-1 strain Ab4p, and zebra-borne EHV-1, was investigated and compared to assess viral kinetics and identify the progression and severity of neuropathological findings. Hamsters were inoculated with EHV-9, EHV-1 strain Ab4p, and zebra-borne EHV-1 via the nasal route and euthanized at 24, 48, 72, 96, 120, 144, and 168 hours postinoculation (HPI). The inoculated hamsters had mild to severe neurological signs at 60 to 72, 96, and 120 HPI, and the mortality rate was 75%, 0%, and 0% for animals inoculated with EHV-9, EHV-1 strain Ab4p, and zebra-borne EHV-1 viruses, respectively. Inoculated hamsters had varying degrees of rhinitis and lymphoplasmacytic meningoencephalitis, as well as differences in the severity and distribution of cerebral lesions. Furthermore, the cellular distribution of viral antigen depended on the inoculated virus. Neuronal necrosis was widely detected in animals inoculated with EHV-9, while marked perivascular cuffs of infiltrating inflammatory cells and gliosis were detected in animals inoculated with EHV-1 strain Ab4p and zebra-borne EHV-1. In the present study, 3 viruses belonging to the herpesvirus family induced encephalitis after initial propagation in the nasal cavity. These viruses might travel to the brain via the olfactory pathway and/or trigeminal nerve, showing different distributions and severities of neuropathological changes.

Ochratoxin A (OTA) formation prevention is not usually available so counteracting strategies are urgent. This study investigated the toxic effects of OTA on Nile tilapia with trials to neutralize these effects by using some feed additives. Supplements used as one percent (Garlen Extra4, Nigella sativa, Garlen Extra4 plus Nigella sativa and a commercial mycotoxins binder, Fero Bind Pro) were added to 500 ppb/kg OTA-contaminated diet. Fish were fed at 3% body weight per day for 10 weeks. The clinical signs recorded in OTA – intoxicated positive control group were sluggish swimming and off food before death with reduction in survivability (53%) and growth performance. Several post-mortem lesions were in liver, kidneys and spleen. Serum levels of ALT, AST, creatinine and urea were significantly increased with reduction in total protein, albumin and globulin in ochratoxicated fish group compared to the negative control group. Concerning the pathological changes that have been noticed in ochratoxin treated fish were almost completely alleviated in examined tissues of fish that were fed on diet with Garlen Extra4 plus Nigella sativa or Nigella sativa and partially in fish that were fed on diet with Garlen Extra4 or Fero Bind Pro, respectively. Best detoxifying results were obtained by using 30 g/kg Nigella sativa plus 0.1 g/kg Garlen Extra4 followed by Nigella sativa, Garlen Extra4 then Fero Bind Pro as a commercial Mycotoxins binder. It could be concluded that inclusions of 30 g/kg Nigella sativa plus 0.1 g/kg Garlen Extra4 in Oreochromis niloticus (O. niloticus) diets could partially reduce OTA toxic effects. Keywords: Detoxifying agents, Feed additives, Garlen, Mycotoxins binder, Nigella sativa

Burullus lagoon is the second largest lake in Egypt. However, there has never been a comprehensive survey which studied nineteen potentially toxic elements in sediments and plants and evaluated the associated potential risk. Thus, we aimed to study the total and potentially available content of As, Al, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Sb, Se, Sn, Tl, V, and Zn in the sediments and common reed (Phragmites australis) at thirty two sites along the entire lagoon and connected drains. Contamination Factor (CF), Pollution Load Index (PLI), Geo-accumulation Index (Igeo), and Enrichment Factor (EF) were calculated to assess the grade of contamination. Element accumulation factor (AF) and bio-concentration ratio (BCR) were also calculated. Aluminum showed the highest median (mg kg−1) total content (41,200), followed by Fe (30,300), Mn (704.7), V (82.0), Zn (75.5), Cr (51.2), Cu (47.8), Ni (44.3), As (31.9), Tl (24.6), Co (21.4), Se (20.3), Sb (17.6), Sn (15.6), Mo (11.3), and Hg (16.6 μg kg−1). Values of the EF, CF, and Igeo showed that the sediments were heavily contaminated with As, Sb, Se, Tl, Mo, Sn, Co, Ni, and Cu. The drained sediment had significantly higher values of total and potentially available element content than the lagoon sediments. Sediments of the middle and western area showed significantly higher contents of total and available elements than the eastern section. The BCR and AF values indicate that the studied plant is efficient in taking up high amounts of Zn, Fe, As, Sn, Tl, Ni, Mo, Mn; then Co, Cu, and V. The results exhibit a dramatic contamination at certain sites of the lagoon, and the studied PTEs have a predominant role in contamination-related ecological risk. Further investigations concerning redox-induced mobilization of PTEs in sediments, the risk of fish contamination and the potential health hazards are highly recommended.

The aim of the present study was to determine whether Moringa oleifera (MOR) leaf powder, have a modulatory effect against gentamicin (GEN)-induced nephrotoxicity in chickens or not. Twenty laying hens were equally divided into four groups: First group (CON) received standard basal diet only. Second group (GEN) was injected with GEN (50mg/kg body weight, intramuscularly (i.m.), twice daily, for 5 consecutive days). Third group (GEN+MOR) was supplemented with MOR leaf powder (10gm/kg diet) daily for 12 consecutive days and injected with GEN (50mg/kg body weight, i.m., twice daily, for 5 consecutive days) starting from the day 8. Fourth group (MOR) was supplemented with MOR leaf powder (10gm/kg diet) daily for 12 consecutive days. GEN administration increased serum creatinine (Cr), blood urea nitrogen (BUN), and malonaldehyde (MDA) and decreased total protein, albumin, reduced glutathione (GSH) content as well as superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities indicating nephrotoxicity. Furthermore, histopathological findings revealed marked tubular vacuolar degeneration, focal interstitial mononuclear cells infiltrations, congestion, focal cystic change and tubular necrosis in GEN-treated kidneys. However, MOR leaf powder supplementation with GEN administration caused significant decrease of serum Cr, BUN, and MDA and increase of total protein, albumin, GSH, SOD, GPx, and CAT compared with GEN alone. Moreover, GEN+MOR treated group showed slight focal interstitial mononuclear cells infiltrations in renal tissues. These findings suggested that dietary supplementation of MOR leaf powder may prevent or reduce the GEN-induced nephrotoxicity.

Pregnant rats were infected experimentally with equine herpesvirus (EHV)-9, a new neurotropic equine herpesvirus serologically similar to EHV-1, during the first and third trimesters. The inoculated dams had mild to severe neurological signs and gave birth to dead fetuses or undersized pups. Rats inoculated during the first and last trimesters had varying degrees of encephalitis as well as abnormalities of the placentas in the form of marked dilation of maternal blood sinusoids and varying degrees of atrophy and necrosis of the trophoblast cells of the labyrinth, the spongiotrophoblasts and the giant cell layer. Virus antigen was detected by immunohistochemistry in the brain and the trophoblast cells of labyrinth, the spongiotrophoblasts and giant cell layer of the placenta in rats inoculated during the first trimester. Virus antigen was detected in fetuses from rats inoculated in the first and last trimesters. Virus DNA was amplified by polymerase chain reaction from the placenta and fetuses of inoculated rats. EHV-9 may induce fetal death and abortion in pregnant dams, possibly caused by direct EHV-9 infection of the placenta and/or fetus as well as the secondary effect of vascular injury.

This study was planned to clarify the biochemical effects of the ginger on CCl4-induced acute hepatotoxicity and its comparison with garlic and combination between ginger and garlic through the determination of the following parameters in the blood and liver tissues of albino rats liver function test. The effect of ginger and garlic against CCL4 was estimated by using white albino rats (150-200 g), which divided into 5 groups of 15 rats. The result showed that CCl4 increase GGT, AST and ALT enzymatic activities, while rats received ginger and / or garlic, their liver function parameters were restored to normal levels and the best result was that group received ginger and garlic after CCL4 injection.

Aflatoxins, toxic metabolic by-products produced by Aspergillus flavus and A. parasiticus, are unavoidable food contaminant and reducing their toxicity in animals is of great interest. The potential of licorice which exhibits antioxidant and anti-inflammatory properties was evaluated for alleviating the AFB1-induced (Aflatoxin B1) hepatotoxicity in rat. Four experimental groups were used, each comprising 5 rats; control group (G1), licorice-received group (G2) (300mg/kg b.w intragastric daily for 7 days), AFB1-treated group (G3) (5 mg/kg b.w. one dose intrapretonial), and a group given licorice 7 days before AFB1 intoxication (G4) 24 hours later, the animals sacrificed. The activities of Alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST) were measured in rat serum as well as histopathological examination of liver. Results revealed the following: AFB1 significantly elevated the serum ALT, AST and the liver showed diffuse necrosis, bile duct hyperplasia, congestion of the central vein and portal mononuclear cells infiltrations. Rats received licorice before AFB1, showed a significant amelioration in serum enzymes and improvement in liver tissues architecture. In conclusion, licorice was found to be safe and successful agent counteracting the toxicity induced by AFB1 in the liver of rat.

In our experimental study we evaluated the efficacy of human freeze-dried amniotic membrane on healing of corneal wound defect in dogs. A corneal wound was performed by using corneal trephine 8.5 mm in diameter. A total of 14 dogs were divided into two groups of 7 dogs in each: (A) Control group; (B) AM group treated by transplantation of freeze-dried human amniotic membrane in conjunction with a third eyelid flap. Clinical outcome was monitored by evaluation of epiphora, ocular discharge, corneal vascularization, corneal opacity and corneal epithelization at the 7th, 11th, 15th and 30th days post surgery. Fluorescein staining was used to estimate the closure of the corneal wounds. Histopathological examination was performed in two dogs of each group; the thickness of the newly generated cornea was measured by using Image j analysis software. The control group was complicated with severe neovascularization, ocular discharge and corneal opacity. There were excessive thickness in the stroma of control group as reflect of inflammatory reaction. The AM group showed mild degree vascularization which was completely absent by the end of experiment. Corneal epithelialization was completed within 15 days after the transplantation in the AM group. In conclusion, transplantation of human amniotic membrane improves canine corneal epithelial wound healing effectively.

This study aimed to clarify the timing and infectivity of equine herpesvirus 9 (EHV-9) infection in BALB/c-nu/nu mice and their immunocompetent counterpart (BALB/c). Following intranasal inoculation with 105PFU of EHV-9, specimens from 8 mice per group were collected at different times postinoculation (PI) and assessed using histopathology, immunohistochemistry for viral antigen, and quantitative real-time polymerase chain reaction for ORF30 gene expression. In BALB/c-nu/nu mice, EHV-9 antigen was abundant in olfactory epithelia of all inoculated animals, and in the olfactory bulb of 1 animal. In contrast, only 1 BALB/c mouse per time point had rhinitis, with mild to moderate immunopositivity starting from 12 to 48 h PI, followed by a gradual virus clearance at 72 h PI. Statistically, significant differences were noted in the immunohistochemistry reactions between the 2 mouse strains, indicating that BALB/c-nu/nu is more susceptible to infection. Relative expression levels of ORF30 gene in olfactory epithelia were significantly different between the 2 groups, with the exception of 12 h PI, when BALB/c-nu/nu animals showed dramatic increases in ORF30 gene expression level until 48 h PI, followed by a decline in expression level until the end of experiment. In contrast, the expression level in brains showed no differences between mouse strain except at 96 h PI. In both strains, the highest messenger RNA expression was detected at 48 h PI, followed by a decline in BALB/c mice, proving a rapid clearance of virus in BALB/c and a gradual slowing down of the increased expression levels in BALB/c-nu/nu.

Summary The pathogenesis and kinetics of oral infection by equine herpesvirus (EHV)-9 were studied in mice and hamsters. After oral inoculation of 105 plaque-forming units (PFU) of virus, 1-week-old suckling hamsters showed varying severity of neurological disease from 72 hours post inoculation (hpi) and all of these animals had died by 96 hpi. Four-week-old ICR mice inoculated orally with 4 × 104 PFU of virus showed no clinical signs, but they developed erosive and ulcerative gastritis from 36 hpi. Varying degrees of encephalitis were seen in infected mice and hamsters, and the hamsters also developed myelitis by 96 hpi. Immunohistochemistry performed on whole body sections of suckling hamsters revealed the kinetics of spread of the virus to the central nervous system. EHV-9 antigen was detected initially in macrophages of the oral and lingual submucosa. At 36 hpi virus antigen was detected in the nerve fibres and pseudounipolar neurons of the trigeminal ganglion and at 96 hpi antigen was present in the myenteric plexuses of the intestine. Virus antigen was also detected in the liver, lungs and heart of affected animals. EHV-9 DNA was detected by polymerase chain reaction in the brain, blood and spinal cord of suckling hamsters at 36, 48 and 96 hpi. These findings show that EHV-9 may spread via the trigeminal nerve when mice and hamsters are inoculated orally with virus.

The kinetics of infection and pathogenicity of equine herpesvirus-9 (EHV-9) was studied in a hamster model. Five-week-old Syrian hamsters and 5-day-old suckling hamsters were inoculated intraperitoneally with 10(5) and 4×10(4) plaque-forming units of EHV-9, respectively. EHV-9 antigens were detected by immunocytochemistry in the peritoneal macrophages, which may be the primary site of virus attachment and propagation at 6h post inoculation (hpi). At 12 hpi, viral antigen was observed in the abdominal nerves and ganglia (mainly the coeliac ganglia). Virus antigen was detected in the dorsal root (spinal) ganglia, in parts of the spinal cord (particularly the mid-lumbar area) and in the myenteric plexuses at 36, 48 and 72 hpi, respectively. At 96 hpi, virus antigen was detected in the most caudal part of the brain. Polymerase chain reaction conducted on samples of the blood, spinal cord and brain revealed EHV-9 DNA in the spinal cord at 36 hpi and in the blood at 48 hpi and for 4 days after this initial detection. It is suggested that after initial propagation in the abdominal macrophages, EHV-9 infected the abdominal ganglia or myenteric plexuses and then travelled to the brain via the peripheral nerves and spinal cord. Examination of other organs also revealed the presence of EHV-9, suggesting that the virus might infect tissues other than those of the nervous system.

The pathogenicity of equine herpesvirus 9 (EHV-9), a neurotropic equine herpesvirus isolated from a herd of Gazella thomsoni, was studied in cattle. Seven calves were inoculated intranasally with 105 and 107 plaque-forming units of the EHV-9 P19. Three animals showed brain lesions consisting of glial reactions and perivascular cuffings in the olfactory bulb and the frontal and temporal lobes. Additionally, the animal that was inoculated with 107 plaque-forming units showed neuronal degeneration and loss, as well as nuclear inclusions compatible with herpesvirus. EHV-9 was isolated from the brain of calf 6 and the lungs of calves 1 and 2. The results suggested that cattle are susceptible to experimental infection with EHV-9 and at risk from natural infection from reservoir hosts.

Equine herpesvirus 9 (EHV-9), the newest member of the equine herpesvirus family, is a highly neurotropic herpesvirus that induces encephalitis in a variety of animals. To access transmission of EHV-9 in the nasal cavity and brain, a suckling hamster model was developed so that precise sagittal sections of nasal and cranial cavities including the brain could be processed, which proved useful in detecting viral transmission as well as extension of pathological lesions. Suckling hamsters were inoculated intranasally with EHV-9, and were sacrificed at 6, 12, 18, 24, 36, 48, and 60 h post inoculation (PI). Sagittal sections of the entire head, including nasal and cranial cavities including the brain, were made to assess viral kinetics and identify the progress of the neuropathological lesions. At 12 to 24 h PI the virus attached to and propagated in the olfactory epithelium, and infected adjacent epithelial cells. At 48 h PI, immunohistochemistry for EHV-9 viral antigen showed that virus had extended from the site of infection into the olfactory bulb and olfactory nerve. These results indicate that EHV-9 rapidly invades the brain via the olfactory route after experimental intranasal infection.

Knowledge of how fish respond to starvation periods could provide a basis for improved nutrition, rearing, disease control and thereby help to optimize Oreochromis niloticus culture. In the current study, the effect of alternative starvation-re-feeding regime was monitored on some growth, hematological and serum biochemical parameters in Nile tilapia fingerlings; followed by checking the role of these feeding schedules in combating the disease challenge. The study was performed in glass aquariums at the aquaculture research unit, Sakha, Kafr El-Sheikh governorate, Egypt using 375 Oreochromis niloticus fingerlings weighing 29±0.5 gram. Fingerlings were subjected to different starvation periods (7, 10, 14, 21 days), followed by 30 days of refeeding on a commercial fish ration (25% protein). Fish growth parameters, hematological and serum biochemical parameters were recorded before starvation, before refeeding and after 30 days of refeeding. The experimental challenge was carried out to determine the effect of different starvation intervals followed by refeeding regimes on the immunity of O. niloticus fingerlings; against bacterial infection with Aeromonas hydrophila through recording mortality rates and the histopathological finding. The collected results revealed that fingerlings can recover all the hematological and serum biochemical parameters values of all food deprived groups close to the normal level of full-fed control fish; concluding that O. niloticus can easily be cultured on suitable economic short-time feeding regime with subsequent refeeding in alternate days without any significant differences in fish size and final production (reducing the food amount and cost required for production cycle). Besides, short-term starvation prior to a bacterial infection followed by subsequent refeeding could promote the defense mechanism of the fish to fight against Aeromonas hydrophila.

Background It was suggested that Equine herpesvirus 9 (EHV-9) could be transmitted to higher non-human primates. Methods Four cynomolgus monkeys (Macaca fascicularis) were inoculated with EHV-9 by the nasal route. Results No abnormalities were observed pathologically, immunohistochemically, and genetically. Conclusions These findings indicate that cynomolgus monkeys are not susceptible to EHV-9.

By using a new member of the neurotropic equine herpesviruses, EHV-9, which induced encephalitis in various species via various routes, an ocular infection model was developed in suckling hamsters. The suckling hamsters were inoculated with EHV-9 via the conjunctival route and were sacrificed after 6, 12, 24, 36, 48, 72, 96, 120, and 144 hours (h) post inoculation (PI). Three horizontal sections of the brains, including the eyes and cranial cavity, were examined histologically to assess the viral kinetics and time-course neuropathological alterations using a panoramic view. At 6 to 24 h PI, there were various degrees of necrosis in the conjunctival epithelial cells, as well as frequent mononuclear cell infiltrations in the lamina propria and the tarsus of the eyelid, and frequent myositis of the eyelid muscles. At 96 h PI, encephalitis was observed in the brainstem at the level of the pons and cerebellum. EHV-9 antigen immunoreactivity was detected in the macrophages circulating in the eyelid and around the fine nerve endings supplying the eyelid, the nerves of the extraocular muscles, and the lacrimal glands from 6 h to 144 h PI. At 96 h PI, the viral antigen immunoreactivity was detected in the brainstem at the level of the pons and cerebellum. These results suggest that EHV-9 invaded the brain via the trigeminal nerve in addition to the abducent, oculomotor, and facial nerves. This conjunctival EHV-9 suckling hamster model may be useful in assessing the neuronal spread of neuropathogenic viruses via the eyes to the brain.

The infectivity and pathology of equine herpesvirus 9 (EHV-9), a new neurotropic equine herpesvirus isolated from gazelles, was studied in hamsters experimentally infected via nasal, ocular, oral, intravenous (IV), or peritoneal routes. Clinically, all animals inoculated by the nasal route and ~25% inoculated by the oral and peritoneal routes showed neurological signs on days 3, 6, and 9 postinoculation (PI), respectively. Neurological signs were not observed in animals administered EHV-9 by the IV and ocular routes. With the exception of animals administered EHV-9 by the IV route, all infected animals had lymphocytic meningoencephalitis. Although there were a number of differences in the severity and distribution of the lesions depending on the route of inoculation, the basic features of lymphocytic meningoencephalitis caused by EHV-9 were common. Lesions consisted of neuronal necrosis, perivascular aggregates of lymphocytes, plasma cells, and neutrophils, gliosis, intranuclear inclusion bodies, and diffuse lymphocytic infiltrates in the meninges. Viral antigen was detected in degenerated neurons in infected animals inoculated by the nasal, ocular, oral, and peritoneal routes. The distribution of EHV-9 antigen was somewhat dependent on inoculation route. There were no microscopic abnormalities or viral antigen in animals treated by the IV route. This study provides new data about experimental EHV-9 infection in hamsters through routes other than the IV route. These results suggest that in the animals infected by the oral, ocular, and peritoneal routes, EHV-9 might travel to the brain through nerves, other than by the olfactory route, after initial propagation at the site of viral entry.

It was suggested that Equine herpesvirus 9 (EHV-9) could be transmitted to higher non-human primates.Four cynomolgus monkeys (Macaca fascicularis) were inoculated with EHV-9 by the nasal route.No abnormalities were observed pathologically, immunohistochemically, and genetically.These findings indicate that cynomolgus monkeys are not susceptible to EHV-9.

The pathogenicity of equine herpesvirus (EHV)-9, a new neurotropic equine herpesvirus isolated from gazelles, was assessed in pregnant rodents (mice and hamsters) following intranasal inoculation. The pregnant female mice and hamsters were inoculated with EHV-9 in the early or late trimesters. The inoculated animals exhibited mild to severe neurological signs and gave birth to dead or undersized fetuses. All three mice and four hamsters inoculated in the first trimester had varying degrees of placental abnormality, characterized by markedly dilated maternal blood sinusoids, atrophy of the trophoblast cells and necrosis of the middle layer of the trophoblast. There was also endometrial blood vessel congestion and necrosis and disorganization of the fetal capillaries in the mice and hamsters inoculated in the last trimester. EHV-9 antigen was detected in the brain of dams and the lungs of the fetuses and in the middle of the trophoblast layer of the placenta in hamsters inoculated in the first trimester. The placental lesions were milder in mice than in the hamsters. The mice and hamsters inoculated in the last trimester had more prominent lesions than the animals inoculated in the first trimester. These results suggest that EHV-9 can cause the death of the fetus or abortion and that these events may be secondary to placental vascular compromise.