Tick-borne diseases caused by Anaplasma species put serious constraints on the health and production of domestic cattle in tropical and sub-tropical regions. After recovering from a primary infection, cattle typically become persistent carriers of pathogens and play a critical role in the epidemiology of the disease, acting as reservoirs of the Anaplasma spp. In this study a duplex PCR assay was used for the simultaneous detection of Anaplasma marginale and Anaplasma phagocytophilum in cattle using two primer pairs targeting msp4 and msp2 genes, respectively. We used this method to analyze DNA preparations derived from 328 blood cattle samples that were collected from 80 farms distributed among Tunisia’s four bioclimatic zones. The prevalence of the A. marginale infection (24.7 %) was significantly higher and more widespread (in all bioclimatic areas) than that of A. phagocytophilum (0.6 %), which was found in a mixed infection with A. marginale. The duplex PCR assay used proved to be a rapid, specific and inexpensive mean for the simultaneous detection of Anaplasma marginale and Anaplasma phagocytophilum in cattle blood. It allowed us to report the identification of A. phagocytophilum for the first time in cattle in Tunisia and confirm the presence of A. marginale in cattle from several geographical areas of the country. Further epidemiological studies undertaken using this assay will help improve the surveillance of the associated diseases in the regions where they are endemic.

The Culex pipiens complex (Diptera: Culicidae) includes the most widespread mosquito species in the world. Members of this complex are the primary enzootic and epidemic vectors of the West Nile virus (genus Flavivirus) in several countries. The two recognized forms of Cx. pipiens (Linnaeus, 1758) - pipiens and molestus - exhibit behavioral and physiological differences. Natural populations of Cx. pipiens were investigated in several sites in Tunisia to evaluate the ecophysiological and molecular characteristics of their forms. The analysis showed the sympatric presence of Cx. pipiens forms and hybrids in all studied sites. Of all the tested larvae of Cx. pipiens, 33.5% were identified as pipiens, 30.8% were identified as molestus, and 35.6% were identified as hybrids. The molestus and hybrid forms were positively correlated with urban habitats and belowground sites while the pipiens form was positively correlated with rural habitats and aboveground sites. Autogeny was expressed in all types of habitats and breeding sites. By contrast with the microsatellite CQ11, the two molecular markers, ace-2 and cytb, did not allow differentiation between the Cx. pipiens forms. Our study shows the ubiquitous distribution and the plasticity of the different forms of Cx. pipiens in a wide range of ecological conditions. It suggests that the behavioral traits assigned to the forms of Cx. pipiens seem to be more flexible than previously assumed. Our analysis also proves that the microsatellite CQ11 remains an efficient tool for distinguishing between Cx. pipiens forms.

Environmental DNA (eDNA) metabarcoding has transformed marine biodiversity monitoring by allowing non-invasive, cost-effective detection of species with high resolution across diverse marine habitats. A systematic literature search was conducted using Google Scholar, Scopus, and the Qatar University Library databases. Relevant peer-reviewed publications were screened and selected based on predefined inclusion criteria to ensure comprehensive coverage of studies. This review synthesizes advances in global and regional eDNA applications, emphasizing the Middle East and North Africa (MENA) region, which faces unique environmental extremes, high endemism, and significant data gaps. eDNA metabarcoding often outperforms traditional methods under comparable sampling effort to traditional surveys in detecting rare, cryptic, and invasive taxa, but technical challenges like incomplete reference databases, primer biases, PCR inhibitors, and inconsistent methodologies limit their effectiveness, particularly in understudied areas such as MENA. Recent developments, including multi-marker approaches, autonomous samplers, and next-generation sequencing, are enhancing detection precision and enabling broader, real-time monitoring. In the MENA region, early studies have revealed eDNA’s potential for habitat distinction, biogeographic research, pollution assessment, and the early discovery of non-indigenous species, although progress is hindered by gaps in reference libraries, infrastructure, and regulation. This review underscores the urgent need for regional collaboration, standardized protocols, and capacity-building. By integrating eDNA with traditional methods and leveraging emerging technologies, the MENA region can establish itself as a leader in marine biomonitoring under extreme environmental conditions, providing actionable insights for conservation and sustainable management of its unique marine ecosystems.