DNA barcoding of Tribolium castaneum (Coleoptera: Tenebrionidae) from selected states in Nigeria based on mitochondrial DNA sequences

Tribolium castaneum also known as red flour beetle is one of the most important pests of stored grain product with a cosmopolitan distribution in Nigeria and all over the world contributing to food spoilage. The aim of this study was to characterize the T. castaneum by morphometric and molecular analyses. Samples of yam flour with evidence of the red flour beetles present inside were obtained from four locations in Kwara, Kogi, Oyo and Ekiti states in Nigeria. Morphological and molecular identifications of T. castaneum were carried out using standard methods. A dissecting microscope was used to identify the beetles and measurements were taken using ImageJ. Genomic DNA was extracted and checked on 1.5% agarose gel to confirm the presence of DNA. Species-specific primers were used to amplify mitochondrial cytochrome oxidase I (COI) gene of T. castaneum and the PCR amplicon size was also checked on 1.5% agarose. Morphometric measurements showed that the highest mean number (33.00±4.24 mm) of T. castaneum larvae observed was recorded on day 61 in Ilorin and the lowest was in Iwo, Osun state (4.00±0.00 mm) on the same day. The mean of the total body length of larvae from sampling sites was (1.31±0.37 mm) with minimum and (1.63±1.14 mm) maximum lengths respectively. There was no significant difference (p>0.05) between the mean length of the larvae collected from the study locations. Aligned cytochrome oxidase subunit 1 (COI) sequences of 313bp were analyzed. Phylogenetic analysis inferred by maximum likelihood method showed that the T. castaneum sequences analyzed for this study and sequences obtained from GenBank formed a monophyletic group. The molecular and phylogenetic analyses confirmed the presence of a single species of T. castaneum. The results from this study showed low levels of genetic diversity and variability in the studied T. castaneum populations. The observed genetic similarity in T. castaneum could be due to the fact that they were probably from similar origin when compared with those in the GenBank database. However, further studies are needed with more samples to characterize T. castaneum species from stored food grains across Nigeria

Genetic Diversity of West African Honey Bee (Apis ‎mellifera adansonii Latreille, 1804) from Rural and Urban ‎Areas of ‎Kwara State, North-Central Nigeria

Over one third of the world’s crops– including fruits, vegetables, nuts, spices, and oilseed–‎require insect pollination, and human reliance on ‎pollination services by honey bees (Apis ‎mellifera) to promote these crops continues to rise due to increasing demands from growing ‎human ‎populations. Identifying the effects of urbanization on genetic diversity on this ‎pollinating insect is important in the field of bioscience. This study aimed to investigate genetic diversity of A. mellifera in Kwara State, Nigeria, using the random amplified polymorphic DNA (RAPD) marker. ‎Thirty honey bees ‎were simultaneously collected from both rural and urban regions in ‎Kwara state, Nigeria. Samples were morphologically identified using ‎standard methods, ‎genomic DNA isolated and amplified using five RAPD primers. Data collected were ‎analysed using PyElph, ‎ARLEQUIN, and GeneAlEx version 6.501 software. The results ‎showed that the DNA fragment sizes produced per primer varied from 200 to ‎‎3000 bp. Percentages of polymorphic loci amplified by each primer varied from 17.33 to 33.33%. ‎Analysis of unbiased Nei genetic ‎distance values showed that Agbede (rural) and Adewole ‎‎(urban) showed the highest value of unbiased genetic distance (0.073), while ‎Amoyo ‎‎(rural) to Idofian (urban) exhibited the lowest value (0.027). Dendrogram analysis revealed ‎genetically close relationships among the sampled ‎A. mellifera‎ populations. The low level of genetic ‎polymorphisms observed among the honey bee populations in the two ‎regions ‎indicated that there is genetic relatedness among them. This study concluded that RAPD ‎marker is a useful method for ‎understanding population genetic structure of the African honey ‎bees. These results can be used as baseline information for future genetic ‎diversity ‎assessment of honey bees in Nigeria with larger samples. It is therefore recommended that ‎there is a need to safeguard the genetic ‎diversity of A. mellifera‎ to prevent extinction or ‎gradual loss of diversity‎‎‎.