Insight into the Toxicological and Pathophysiological Effects of Moroccan Vipers' Venom: Assessing the Efficacy of Commercial Antivenom for Neutralization.

Morocco is one of the richest countries in biodiversity in the Mediterranean region, especially in its ophidian fauna. In total, there are eight species of venomous snakes, with seven belonging to the Viperidae family, responsible for 67.2% of severe envenomation cases in the country. Cerastes cerastes, Daboia mauritanica and Bitis arietans are considered among the most venomous vipers whose bites cause high levels of morbidity, disability or mortality. Despite their wide distribution in the kingdom, the incidence of these snakebites remains poorly understood and largely underestimated. Moreover, intraspecific variations in the venom composition significantly affect the effectiveness of antivenoms. Due to the unavailability of locally produced antivenoms, we evaluated the efficacy of Inoserp-MENA, the only available antivenom in Morocco, against C. cerastes, D. mauritanica and B. arietans. First, we conducted a comprehensive characterization of these venoms, including an LD50 test to examine their toxicity and SDS-PAGE as a technique to analyze the enzymes responsible for biological activities, such as hemorrhagic and edematous activities and myotoxicity, which generate physiopathological effects in the skin, paws and muscles of envenomed mice. Then, we assessed the ability of Inoserp-MENA antivenom to neutralize the toxic activities of Moroccan vipers. Our results indicate that the venom of C. cerastes, D. mauritanica and B. arietans are toxic, causing severe alterations such as edema, myotoxicity, myonecrosis and significant hemorrhages with the formation of hemorrhagic foci. C. cerastes venom is more dangerous in terms of lethality and hemorrhages, while B. arietans venom is more edematous. The effects of C. cerastes venom were effectively neutralized, but Inoserp-MENA antivenom failed to protect mice against the toxic effects induced by B. arietans and D. mauritanica venom. The study reveals alarming shortcomings in the effectiveness of the current commercially available antivenom's dosage and neutralization capabilities, highlighting the urgent need to develop a region-specific viper envenomation therapy.

Assessing the Efficacy of Monovalent and Commercialized Antivenoms for Neutralizing Moroccan Cobra Naja haje Venom: A Comparative Study.

In Morocco, eight species of venomous snakes belonging to the Viperidae and Elapidae families are responsible for severe envenomation cases. The species from the Elapidae family is only represented by the medically relevant cobra Naja haje, which is widely distributed in North Africa. However, there is little information on the systemic effects of Moroccan cobra venom on vital organs due to regional variations. It has been demonstrated that the venom of Naja haje from Egypt causes hemorrhage, while the venom of the Moroccan cobra is neurotoxic and devoid of systemic bleeding. This variability is known to significantly influence treatment efficacy against Naja haje cobra bites in the Middle East. In this study, we examined the pathophysiological mechanisms responsible for the lethality induced by Naja haje venom, as well as the evaluation of the neutralizing capacity of two antivenoms; the monospecific antivenom made for Naja haje only and the antivenom marketed in the Middle East and North Africa. We first determined the toxicity of Naja haje venom by LD50 test, then compared the neutralizing capacity of the two antivenoms studied by determining the ED50. We also performed histological analysis on Swiss mice envenomed and treated with these antivenoms to observe signs of cobra venom envenomation and the degree of reduction of induced systemic alterations. The results showed significant differences between both antivenoms in terms of neutralization. The monospecific antivenom was four times more effective than the marketed antivenom. These results were confirmed by a histological study, which showed that monospecific antivenoms neutralized severe signs of mortality, such as congestion of blood vessels in the heart and kidneys, pulmonary and renal edema, cytoplasmic vacuolization of hepatocytes in the liver, and infiltration of inflammatory cells in the brain and spleen. However, the polyvalent antivenom failed to protect all severe lesions induced by Naja haje venom in mice. These findings highlight the negative impact of geographic variation on the effectiveness of conventional antivenom therapy and confirm the need for a specific Naja haje antivenom for the effective treatment of cobra envenomation in Morocco.

Comparative Proteomic Analysis of the Venoms from the Most Dangerous Scorpions in Morocco: Androctonus mauritanicus and Buthus occitanus.

Morocco is known to harbor two of the world's most dangerous scorpion species: the black Androctonus mauritanicus (Am) and the yellow Buthus occitanus (Bo), responsible for 83% and 14% of severe envenomation cases, respectively. Scorpion venom is a mixture of biological molecules of variable structures and activities, most of which are proteins of low molecular weights referred to as toxins. In addition to toxins, scorpion venoms also contain biogenic amines, polyamines, and enzymes. With the aim of investigating the composition of the Am and Bo venoms, we conducted an analysis of the venoms by mass spectrometry (ESI-MS) after separation by reversed-phase HPLC chromatography. Results from a total of 19 fractions obtained for the Am venom versus 22 fractions for the Bo venom allowed the identification of approximately 410 and 252 molecular masses, respectively. In both venoms, the most abundant toxins were found to range between 2-5 kDa and 6-8 kDa. This proteomic analysis not only allowed the drawing of an extensive mass fingerprint of the Androctonus mauritanicus and Buthus occitanus venoms but also provided a better insight into the nature of their toxins.

Antibiotic Susceptibility of ß-Glucuronidase-Positive Escherichia coli Isolated from Poultry Products in Morocco.

Poultry products are one of the main foodstuffs consumed in Morocco. The deterioration of their hygienic quality and the emergence of antibiotic resistance are the major public health problems. This study was carried out to determine the level of emergence of antibiotic resistance. For that, between May 2021 and June 2022, poultry products (e.g., minced meat, sausage, and meat) were collected aseptically in sterile bags from retail stores in different cities of Morocco, labeled, and transported in chilled conditions to the laboratory. The bacterial analysis was performed within 24 hours. Isolation and identification of Escherichia coli were performed according to the Moroccan standard NM ISO 16649 2018. Antimicrobial susceptibility determination of 23 antibiotics was tested using the Kirby-Bauer disk diffusion method. The results showed a high level of antimicrobial resistance to amoxicillin (58%), amoxicillin-clavulanic acid (54%), piperacillin (63%), trimethoprim (32%), nalidixic acid (46%), and ofloxacin (43%). 58 isolates (60%) were positive for beta-lactamase class A (penicillinase) test detection, and 2 isolates (3%) produced extended-spectrum beta-lactamases. The high level of resistance demonstrated in this study should alert health organizations in the country. An establishment of surveillance programs to control the use and the administration of antibiotics in the poultry field and initiation of reliable methods to follow up on the latest updates on the emergence of bacterial resistance is a necessity.

Bacteriological Quality of Table Eggs in Moroccan Formal and Informal Sector.

Eggs constitute an important part of the Moroccan diet. However, contaminated eggs can cause a serious public health problem if consumed undercooked, uncooked, or used in unpasteurized egg foodstuffs. This study was carried out to evaluate the microbial contents of eggs according to their sales sector in Morocco. For that, a total of 1770 eggs were collected from January to September 2021 from formal markets (refrigerated eggs from large shopping centers) and informal markets (eggs at ambient temperature from ambulatory sellers, street vendors, kiosks, and neighborhood stores) and transferred to the Avian Pathology Unit at Hassan II Agronomic and Veterinary Institute. The eggshells and their contents were tested separately; swabs of eggshells were used to inoculate Mac-Conkey agar, while the egg contents were cultured on Mac-Conkey and Mannitol salt agar, then standard microbiological tests were performed to identify the isolated organisms. The results showed that informal eggs were more contaminated (87%) than formal eggs (48) (p < 0.05). The bacteria isolated from the eggshells (informal and formal) were Enterobacter agglomerans (59% and 21%), Klebsiella spp. (24% and 4%), Enterobacter cloacae (17% and 8%), E. coli (9% and 1%), Serratia spp. (9% and1%), Pseudomonas aeruginosa (9% and 1%), Shigella spp. (5% and 0%), Salmonella enteritidis (0% and 2%), Proteus spp. (4% and 0%), Enterobacter sakazakii (2% and 0%), Rahnella aquatilis (1% and 0%), and Staphylococcus aureus (0% and 1%). For the egg-contents, the detected bacteria (informal and formal) were Enterobacter agglomerans (14% and 28%), Klebsiella spp. (7% and 6%), Staphylococcus aureus (6% and 1%), Enterobacter cloacae (4% and 4%), E. coli (4%, 1%), Shigella spp. (4%, 0%), Acinetobacter baumannii (3% and 1%), Salmonella enteritidis (2% and 0%), Serratia spp. (1% and 6%), Proteus spp. (1% and 3%), and Enterobacter sakazakii (1% and 0%). We conclude that eggs might be contaminated with several bacteria and can constitute a public health threat in Morocco.

A wheat lipid transfer protein (TdLTP4) promotes tolerance to abiotic and biotic stress in Arabidopsis thaliana.

Lipid transfer proteins (LTPs) are members of the family of pathogenesis-related proteins (PR-14) that are believed to be involved in plant defense responses. In this study, we report the isolation and characterization of a novel gene TdLTP4 encoding an LTP protein from durum wheat [Triticum turgidum L. subsp. Durum Desf.]. Molecular Phylogeny analyses of wheat TdLTP4 gene showed a high identity to other plant LTPs. Predicted three-dimensional structural model revealed the presence of six helices and nine loop turns. Expression analysis in two local durum wheat varieties with marked differences in salt and drought tolerance, revealed a higher transcript accumulation of TdLTP4 under different stress conditions in the tolerant variety, compared to the sensitive one. The overexpression of TdLTP4 in Arabidopsis resulted in a promoted plant growth under various stress conditions including NaCl, ABA, JA and H2O2 treatments. Moreover, the LTP-overexpressing lines exhibit less sensitivity to jasmonate than wild-type plants. Furthermore, detached leaves from transgenic Arabidopsis expressing TdLTP4 gene showed enhanced fungal resistance against Alternaria solani and Botrytis cinerea. Together, these data provide the evidence for the involvement of TdLTP4 gene in the tolerance to both abiotic and biotic stresses in crop plants.

Evidence for non-disomic inheritance in a Citrus interspecific tetraploid somatic hybrid between C. reticulata and C. limon using SSR markers and cytogenetic analysis.

Artificial tetraploid somatic hybrids have been developed for sterile triploid citrus breeding by sexual hybridization between diploid and tetraploid somatic hybrids. The genetic structure of diploid gametes produced by tetraploid genotypes depends on the mode of chromosome association at meiosis. In order to evaluate tetraploid inheritance in a tetraploid interspecific somatic hybrid between mandarin and lemon, we performed segregation studies using cytogenetic and single sequence repeat molecular markers. Cytogenetic analysis of meiosis in the somatic hybrid revealed 11% tetravalents and 76% bivalents. Inheritance of the tetraploid hybrid was analyzed by genotyping the triploid progeny derived from a cross between a diploid pummelo and the tetraploid somatic hybrid, in order to derive genotypes of the meiospores produced by the tetraploid. A likelihood-based approach was used to distinguish between disomic, tetrasomic, and intermediate inheritance models and to estimate the double reduction rate. In agreement with expectations based the cytogenetic data, marker segregation was largely compatible with tetrasomic and inheritance intermediate between disomic and tetrasomic, with some evidence for preferential pairing of homoeologous chromosomes. This has important implications for the design of breeding programs that involve tetraploid hybrids, and underscores the need to consider inheritance models that are intermediate between disomic and tetrasomic.