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The present study was conducted in order to isolate, identify and characterize fowl aviadenovirus associated with inclusion body hepatitis (IBH) in three poultry farms (two of broiler chickens and one of breeder broiler chickens) in Morocco during 2015. Liver samples collected from affected three poultry farms were examined by histopathological examination. Tissue samples showing necrosis of hepatocytes associated with basophilic intranuclear inclusion bodies were homogenized and submitted to FAdV isolation in chicken embryo fibroblast (CEF) cell cultures and in SPF embryonated eggs. The cytopathic effect (CPE) was observed in the second passage with swelling and rounding of infected cells. The inoculated embryos were hemorrhagic and showed hepatitis with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The presence of the virus was confirmed by conventional polymerase chain reaction based on hexon gene from all investigated samples. Moreover, phylogenetic analysis of the hexon gene revealed that FAdVs isolated from different affected poultry belonged to FAdV 11 serotype of the D genotype group. This work is the first isolation in cell culture and SPF embryonated eggs of FAdV from Moroccan broilers and breeder broiler chickens with IBH.

Outbreaks of inclusion body hepatitis have emerged in Morocco since 2013 and has resulted in significant economic losses to poultry farms. Three isolates of the causative virus, Fowl adenonovirus (FAdV)were characterized from chickens with IBH, but their pathogenicity has never been investigated. In this work, the pathogenicity of an isolate FAdV 11 (MOR300315 strain) was evaluated by inoculating a group of 40 SPF chickens at 3 days of age by oral route. A group of 40 chicks injected with phosphate-buffered saline solution was used as a control group. The infected chickens showed decreased weight gain from 3dpi. Necropsy displayed pallor and enlargement in liver, swelling and slight hemorrhage in kidney and spleen at 6 dpi. Histopathological changes were mainly characterized by severe and extensive hepatic necrosis associated with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The FAdV was reisolated in chicken embryo fibroblast cell culture from liver tissue homogenate of infected chicken from 3 to 6 dpi. Viral DNA was detected by PCR in liver, kidney, spleen and cloacal swabs from 3 to 13 dpi. Antibody response against inoculated FAdV was appeared from 9 dpi. These results confirmed that the FAdV 11 strain is pathogenic in chicken. This study is the first experimental infection of FAdV 11 in chicken in Morocco, which increase our understanding of its pathogenicity in chickens and indicate that preventive measures against FAdV infection in poultry farms should be implemented in Morocco.

Four aromatic and therapeutic plants, Thymus vulgaris, Rosmarinus officinalis, Pimpinella anisum, and Foeniculum vulgare, were examined in this comparative study. The objectives were to assess its phytochemical composition; polyphenol, flavonoid, and tannin content; antioxidant and antibacterial activity; bioactive molecule identification; and critical trace element quantification. Its aqueous and organic extracts were examined, focusing on ethanolic extracts. The ethanolic extract’s ability to neutralize free radicals was validated by phytochemical studies and antioxidant tests, underscoring their role in preventing oxidative stress. An Ultra-Performance Liquid Chromatography—High-Resolution Mass Spectrometry Orbitrap Exploris 120 (UPLC–HRMS Orbitrap) was used to identify the bioactive chemicals, and the results showed a variety of compounds having antibacterial and antioxidant properties. The important trace elements found in these plants were also measured using a Graphite Furnace-Atomic Absorption Spectrometer (GF-AAS). These components are essential to the biological characteristics of the plants, especially their antioxidant and antibacterial capacities. Among the aqueous extracts, it was observed that Rosmarinus officinalis and Foeniculum vulgare exhibited a MIC of 3.91 µg/mL against Staphylococcus. Additionally, R. officinalis also demonstrated a MIC of 3.91 µg/mL against Escherichia coli. All of the data were interpreted and shown using principal component analysis. The results were grouped and explained using this statistical method, which revealed a strong association between the abundance of antibacterial and antioxidant chemicals in the four plants under investigation.

The phytochemical profiles, antioxidant capacities, mineral composition, and antibacterial activities of Zingiber officinale (Z. officinal) and Piper nigrum (P. nigrum) were explored through aqueous, ethanolic, and methanolic extractions. The extracts were analyzed for polyphenols, flavonoids, and tannins, and their antioxidant potential was assessed using the DPPH assay. UPLC-HRMS identified major bioactive compounds, including 6-gingerol and shogaol in Z. officinale, and piperine and piperlonguminine in P. nigrum. Mineral analysis showed that P. nigrum was particularly rich in essential elements, including calcium (Ca), magnesium (Mg), and iron (Fe). In antibacterial testing, P. nigrum demonstrated wider zones of inhibition against E. coli, whereas Z. officinale was more active at lower concentrations, showing MICs as low as 3.91 µg/mL against Salmonella and S. aureus. PCA analysis revealed strong correlations between phenolic content and biological effects. These results underscore the potential of both spices as effective natural agents for use in food preservation and health-promoting applications.

The concerns regarding Fowl Adenoviruses have gained significance in the poultry industry due to their association with various diseases, including Inclusion Body Hepatitis, Hepatitis-Hydropericardium Syndrome, and Adenoviral Gizzard Erosion (AGE). AGE is an emerging disease reported in several countries, particularly in Asia and Europe, causing significant economic losses in the poultry industry. In 2001, Fowl Adenovirus Serotype 1 was identified as the etiological agent of AGE in Japan. Since then, it has been spreading to other countries due to its transmission mode. Although Adenoviral Gizzard Erosion has been mostly described in broilers, it has also been observed in layers and pullets. This review offers a comprehensive analysis of Fowl Adenovirus Serotype 1, encompassing various key aspects of the virus. We also examine the pathogenesis and epidemiology of the virus, providing an overview of its distribution and prevalence in avian populations worldwide. Highlighting the most recent developments in serological and molecular techniques for virus detection, quantification, and genotyping and comparing them to conventional tests, this review aims to contribute to the understanding of the diagnostic workflow for this virus. Lastly, this review sheds light on some vaccine strategies to prevent Adenoviral Gizzard Erosion.

Backyard poultry farming is an important tool for poverty alleviation and food security in rural areas of Morocco. A descriptive epidemiologic survey was conducted in 286 backyard poultry flocks from the provinces of Khemisset and Skhirat-Temara to gain baseline data on the current status of backyard poultry flocks in Morocco as well as its potential implications on the transmission and spread of avian diseases. The findings indicated that 88.8% of flocks were raised in a mixed confinement system, with an average flock size of 30 birds (range 1–352). Chickens accounted for 83% of the overall reported birds. More than two-thirds of respondents (69%) kept chickens only, while the remaining flocks raising multiple bird species in total promiscuity. Diseases were the highest cause of mortality (84.7%), followed by predation (15.3%). According to 56.1% of the owners, respiratory symptoms were among the major disease signs reported, besides ectoparasite infestation. Flock health management revealed a lack of preventive vaccination, lack of veterinary consulting, lack of biosecurity practices, and irrational self-medication of diseased birds using antibiotics, pesticides, and hazardous chemicals that could be a significant health risk for consumers. The need for an outreach program about disease prevention and biosecurity practices, along with prophylactic campaigns, should be emphasized to further mitigate the risks of backyard poultry flocks on the commercial sector and public health.

Fowl adenoviruses (FAdVs) are widespread viruses in poultry populations, responsible for several severe diseases, including Inclusion Body Hepatitis (IBH), Adenoviral Gizzard Erosion (AGE), and Hepatitis-Hydropericardium Syndrome (HHP). These diseases have been associated with significant economic and health impacts on poultry industries. Accurate detection and genotyping play a key role in the diagnosis of these infections, as different FAdV genotypes are associated with distinct disease syndromes and epidemiological patterns. In this study, we aimed to evaluate the clinical, analytical, and genotyping performance of the Hex L1 PCR combined with High-Resolution Melting (HRM) Curve analysis for investigating recent IBH and AGE outbreaks in Morocco. The study involved 26 clinical samples collected from broiler and layer poultry farms suspected with IBH or AGE. These samples were amplified using conventional PCR, real-time PCR/52 K test, and the Hex L1 PCR/HRM test. Field samples were also sequenced and compared with HRM curve analysis results to validate the genotyping accuracy of the Hex L1 PCR/HRM method. Phylogenetic analysis of the sequenced samples revealed several FAdV genotypes, including FAdV-11 and FAdV-8b in IBH cases, and FAdV-1 and FAdV-8a in AGE cases, highlighting the genetic diversity of circulating strains. The Hex L1 PCR/HRM method successfully amplified all 12 FAdV serotypes, demonstrating excellent reproducibility and repeatability, with coefficients of variation ranging from 0.19% to 1.82%. Moreover, this method showed a strong correlation with the real-time PCR/52 K test, achieving a high correlation coefficient of 0.9077. The HRM curve analysis accurately genotyped all the field samples, with results consistent with sequencing outcomes. In conclusion, this method provides a fast, sensitive, and reliable alternative for FAdV detection and genotyping. It enables universal detection, quantification, and genotyping in a single step, overcoming the limitations of traditional techniques, making it an ideal tool for sample screening, while sequencing validation is necessary for confirmation.

Fowl adenoviruses (FAdV) are prevalent in chickens worldwide, responsible for several poultry diseases, including inclusion body hepatitis (IBH), hepatitis-hydropericardium syndrome (HHS), and gizzard erosion (GE), which result in significant economic losses in the poultry industry. Consequently, detection and efficient identification of FAdV serotypes are becoming extremely urgent to monitor outbreaks and develop vaccination strategies. Conventional PCR (cPCR) tests, combined with Restriction Fragment Length Polymorphism (RFLP) or sequencing, were developed for FAdV diagnosis. Although these molecular tests have considerably improved the accuracy of FAdV diagnosis compared with conventional methods, certain drawbacks remain unresolved, including lack of sensitivity and post-PCR analysis. Subsequently, advanced molecular technologies such as real-time PCR (qPCR), Loop Isothermal Amplification (LAMP), Cross-Priming Amplification (CPA), Recombinase Polymerase Amplification (RPA), Digital Droplet Polymerase Chain Reaction (ddPCR), Dot Blot Assay Combined with cPCR, Nanoparticle-Assisted PCR (nano-PCR), PCR-Refractory Quantitative Amplification (ARMS-qPCR), CRISPR/Cas13a Technology, and High-Resolution Melting Curve (HRM), have been developed to improve FAdV diagnosis.

Canine leishmaniasis (CanL) is a zoonotic vector-borne disease that is endemic in the Mediterranean Basin including Morocco. Dogs play a major epidemiological role in this zoonosis as reservoir hosts. This study investigated the clinical manifestations of CanL in dogs naturally infected with Leishmania infantum. A total of 96 dogs presented to the Small Animal Clinic of the Hassan II Agronomy and Veterinary Institute (IAV Hassan II) of Rabat, Morocco, and were tested by RT-PCR and/or serology. Among them, 32 (33.3%) were positive to Leishmania infantum infection. The majority of the positive dogs (93.7%) came from urban areas. Most of them were male (62.5%) and purebreds (65.6%), were aged between 3 and 7 years (71.8%), and had outside activities (guarding, hunting, livestock guarding, and service activities) (71.8%) and all of them were living exclusively outdoor or had free access to the outdoor environment. Lymphadenomegaly (81.2%), dermatological disorders (65.6%) (mostly exfoliative dermatitis), weight loss (59.3%), exercise intolerance (56.2%), anorexia (28.1%), hyporexia (15.6%), and ocular lesions (28.1%) were the most frequent clinical signs and complaints recorded. Anemia and hyperproteinemia due to hyperglobulinemia were observed in 68.7% and 72.7% of the cases, respectively. These results suggest that CanL leads to various nonspecific clinical signs as described previously, making the diagnosis challenging. Since CanL is endemic in Morocco, it should be recommended to systematically test dogs displaying clinical signs compatible with this disease and to regularly screen asymptomatic at-risk dogs. It is also crucial to educate dog owners about the zoonotic aspect of the disease and to encourage intersectorial collaboration following the “One Health” concept, in order to contribute to a more effective control/prevention of human and canine leishmaniasis.

Since 2016, low pathogenic avian influenza virus (LPAIV) H9N2 became a major issue for poultry production in Morocco. Even though the agent was classified as low pathogenic, AI H9N2 cause significant economic losses, particularly during co-infections. Experimentally, it has been difficult to reproduce the clinical picture without appealing other viral or bacterial pathogens. Our study was carried out to evaluate a new challenge model using cold stress in commercial broilers infected with two Moroccan H9N2 viruses isolated in 2016 and 2022. One hundred twenty day-old chicks were divided into four groups: A, B, and C exposed to cold stress, and D was kept as negative control. At 21 days of age, Groups A and B were challenged by oculo-nasal route with 107 EID50 of H9N2 strains, isolated respectively during 2016 and 2022. Meanwhile, chicks of group C were exposed to only cold stress. The assessment of body weight gain, clinical signs, lesions, mortality, and oropharyngeal viral shedding was monitored for 15 days post-challenge. Results showed that cold stress exacerbated H9N2 clinical signs, allowing us to establish a scoring system and to validate the challenge model without co-infections. Gross and microscopic lesions, induced by the virus primarily in the respiratory tract, peaked at 5 dpi and significantly decreased at 15 dpi. Group B harbored the highest viral loads with viral shedding persisting beyond 11 dpi in both groups. This study demonstrates a clear clinical differenceamong the two isolates; A/chicken/Morocco/178-2/2022(H9N2) showed a significant increase in virulence compared to the firstly isolate A/chicken/Morocco/SF1/2016(H9N2).The novel H9N2 challenge model using cold stress will contribute to a better understanding of LPAI pathogenesis and epidemiology and allow for research closer to the field.