Alterations in Histone Methylation States Increased Profusion of Lethal(2)-Essential-for-Life-Like (l(2)elf), Trithorax and Polycomb Genes in Apis mellifera under Heat Stress.

Histone post-translational modifications (PTMs) represent a key mechanism in the thermal adaptation of the honeybee Apis mellifera. In this study, a chromatin immunoprecipitation assay and qPCR were employed to explore the changes in the methylation states of H3K4m2, H3K4m3, H3K27m2 and H3K27m3 associated with l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) ((trx) and PR-set7) and Polycomb (Pc) and (Su(z)12) genes in A. m. jemenitica (tolerant subspecies) and A. m. carnica (susceptible subspecies) in response to heat treatment (42 °C for 1 h). The results revealed significant enrichment fold changes in the methylation/demethylation of most H3K4 and H3K27 marks at all targeted genes. These changes increased the profusion of l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) (trx) and Polycomb (Pc) and Su(z)12 and decreased the profusion of HMT (PR-set7) in both honeybee subspecies. The changes in the methylation enrichment folds of histone methyltransferases (HMTs) ((trx), PR-set) and Polycomb (Pc), Su(z)12 genes demonstrate the well-harmonized coordination of epigenetic gene regulation in response to heat treatment. Compared to the control, the changes in the methylation enrichment folds of H3K4m3 at Polycomb Su(z)12 were about 30× and 100× higher in treated A. m. jemenitica and A.m. carnica, respectively. Similarly, changes in the methylation/demethylation enrichment folds of HMT (trx) and Polycomb (Pc) and Su(z)12 were 2-3× higher in A. m. carnica than in A. m. jemenitica after treatment (42 °C). It is evident that post-translational chromatin modification in both honeybee subspecies can diminish heat stress impact by (I) increasing the transcriptional provision of l2efl associated with survival and (II) increasing the silencing of genes associated with general cellular activities.

Faunistic inventory, identification keys and zoogeographical analysis of the Thysanoptera-Terebrantia of Saudi Arabia, including two new species.

This faunistic study of the Thysanoptera suborder Terebrantia of Saudi Arabia has revealed 79 species in 39 genera and four families. Two families Melanthripidae Bagnall and Stenurothripidae Bagnall, also 25 genera and 51 species, are newly recorded for Saudi Arabia, and Mycterothrips arabicus sp. n. and Scirtothrips erectus sp. n., are described. Males of Thrips lomatus zur Strassan and Odontothrips elbaensis Priesner are described for the first time. Keys to the families, genera and species of Terebrantia of Saudi Arabia are presented, together with a species check list. Zoogeographical analysis indicates that general affinity of the thrips fauna of Saudi Arabia is dominated by Afrotropical (22.6%) and Saharo-Arabian (17.9%) species, followed by Cosmopolitan (16.7%), endemic (11.9%), Palearctic (10.7%), Oriental (9.5%), Mediterranean (7.1%), and traces of Nearctic (2.4%) and Holarctic (1.2%) species.

Expression Levels of Heat-Shock Proteins in Apis mellifera jemenetica and Apis mellifera carnica Foragers in the Desert Climate of Saudi Arabia.

A. m. jemenetica is the indigenous honeybee of the Arabian Peninsula. It is highly adapted to extreme temperatures exceeding 40 °C, yet important molecular aspects of its adaptation are not well documented. In this study we quantify relative expression levels of small- and large-molecular-weight heat-shock proteins (hsp10, hsp28, hsp70, hsp83, hsp90 and hsc70 (mRNAs)) in the thermos-tolerant A. m. jemenetica and thermosusceptible A. m. carnica forager honeybee subspecies under desert (Riyadh) and semi-arid (Baha) summer conditions. The results showed significant day-long higher expression levels of hsp mRNAs in A. m. jemenetica compared to A. m. carnica under the same conditions. In Baha, the expression levels were very modest in both subspecies compared those in Riyadh though the expression levels were higher in A. m. jemenetica. The results also revealed a significant interaction between subspecies, which indicated milder stress conditions in Baha. In conclusion, the higher expression levels of hsp10, hsp28, hsp70ab, hsp83 and hsp90 mRNAs in A. m. jemenetica are key elements in the adaptive nature of A. m. jemenetica to local conditions that enhance its survival and fitness in high summer temperatures.

Linking Histone Methylation States and hsp Transcriptional Regulation in Thermo-Tolerant and Thermo-Susceptible A. mellifera L. Subspecies in Response to Heat Stress.

Genetic and epigenetic responses to environmental cues of worker honeybees mediate hsp synthesis, a key mechanism to tolerate high ambient temperatures in Apis mellifera. In this study, the chromatin immunoprecipitation assay followed by qPCR were used to determine alterations in histone methylation states (H3K27me2, H3K27me3, H3K4me2, and H3K4me3) associated with hsp/hsc/trx in A. m. jemenetica (thermo-tolerant subspecies) and A. m. carnica (thermo-susceptible subspecies) after heat treatment. The results revealed significant changes in enrichment folds of histone methylation states associated with hsp/hsc/trx. Indeed, the enrichment of H3K27me2 decreased strongly in response to heat stress. Changes in histone methylation states were significantly higher in A. m. carnica samples compared to A. m. jemenitica samples. Our study provides a new perception on linking histone post-translational methylation as an epigenetic mechanism of gene regulation with hsp/hsc/trx in A. mellifera subspecies exposed to heat stress.

Assessment of genetic variation in Apis mellifera jemenitica (Hymenoptera: Apidae) based on mitochondrial Cytochrome Oxidase Subunit II and III.

Morphometric and genetic characterization of many Apis mellifera subspecies are well-documented. A. m. jemenetica occurs naturally in Africa and Asia. In this study, genetic variation of mitochondrial Cytochrome Oxidase II (COII) and III (COIII) were analysed in 133 specimens of the endemic honeybee colonies within Saudi Arabia. The COII gene sequence length was 684 bp comprising nine synonymous (1.3%) and two non-synonymous single nucleotide polymorphisms (SNPs) (0.87%). Five variants of COII were not previously documented, one variant (MT755968) showed an extra restriction site when subjected to type II restriction endonuclease from Arthrobacter protophormiae (Apol) or to Haemophilus influenzae Rf (Hinf1). Changes in COII sequence separated samples into three haplogroups. Whereas, COIII gene sequence length was 780 bp, including 18 synonymous and five non-synonymous SNPs. Furthermore, variation in COII sequence was more informative based on restriction profiles and on amino acid changes compared with COIII gene sequence. Variants of COIII showed identical restriction sites when subjected to type II restriction endonuclease from Deinococcus radiophilus (DraI), and revealed high similarity to African subspecies. Results of this study are very useful in understanding genetic diversity and characterization of A. mellifera subspecies.

Assessment of genetic variation in Apis mellifera jemenitica (Hymenoptera: Apidae) using Cytochrome Oxidase I gene sequences.

The Arabian Honeybee Apis mellifera jemenitica is endemic to the Arabian Peninsula. It is highly adapted to temperature extremes and drought dominating the region. In this study, the mitochondrial Cytochrome Oxidase I (COI) was analyzed in 133 specimens of A. m. jemenitica from eight localities along the Red Sea cost of Saudi Arabia. Results revealed 33 synonymous, and 6 non-synonymous mutations within the COI sequences, resulting in change of 4 amino acids. Phylogenetic analysis based on either type of mutations revealed two main haplogroups accounting for 94% of the samples. In total Eighteen new haplotypes were identified and uploaded in the genebank, Fourteen of them are restricted to one/both haplogroups. All haplotypes identified in this study clustered with reference COI sequences of the sub-lineag Z (African Lineage). However one Haplotype (MW428270) represents high COI variability compared to other haplotypes and may resemble different evolutionary sub-lineage. Tajima's Neutrality Test (Ps = 0.025; D = -1.5) indicated population size expansion that took place after selective sweep and/or purifying selection.

The Thripidae subfamily Sericothripinae (Thysanoptera) from Saudi Arabia with a new species of Hydatothrips Karny.

The subfamily Sericothripinae Karny (Thysanoptera: Thripidae) is recorded for the first time from the Saudi Arabian fauna. Four species belonging to two genera, Hydatothrips Karny and Neohydatothrips John, are recognized, and H. bahaensis Rasool, sp. n. is described from Al Baha region (southwestern of Saudi Arabia). Host-plant associations are given for the new species and also N. amygdali Minaei, and the male and larvae of N. amygdali are reported for the first time. An illustrated key to the genera and species is provided.

Mating frequency of Apis mellifera jemenitica under desert conditions of Saudi Arabia.

Queen mating frequency is an important reproductive trait of the western honeybee Apis mellifera. Yet, it demands more attention when investigated under extreme or confined ecosystems. Queen mating frequency of the Yemeni Honeybee A. m. jemenetica was estimated under Saudi Arabia desert conditions, Riyadh (24°71'36″N, 46°67'53″E). Mating of queens took place after 8-13 days from emergence. Duration of mating flight ranged between 26 and 39 min. Subsequently, six microsatellite loci were used to genotype queen's progeny (n = 30 workers/queen). The average number of drone alleles using workers genotypes ranged between 5.83 ± 0.31 and 6.33 ± 1.09. However, effective paternal allele number was extremely low and ranged between 3.35 ± 0.34 and 3.60 ± 0.40. This relatively low mating frequency of the Yemeni honeybee, A. m. jemenetica, might have striking effect on the overall colony survival. Providentially, this relatively low mating frequency does not impact colonial heterozygosity, shown in this study (0.66 ± 0.07-70 ± 0.04), adversely. These results may affect hive survivability and entails distinctive management practices under such conditions.

The complete mitochondrial genome and phylogenetic analysis of the Arabian Honeybee Apis mellifera jemenitica (Insecta: Hymenoptera: Apidae) from Saudi Arabia.

Fourteen mitochondrial genomes from workers of the Arabian Honeybee Apis mellifera jemenitica were determined. Genomes range from 16,352 to 16,445 bp. Each consists of 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and one control region. The mitogenome sequences revealed 753 Variable sites in total, distributed within protein-coding loci (199), ribosomal RNAs genes (117), transfere RNAs genes (48), and non-coding AT-rich region's (389). Phylogenetic analysis with Neighbor-Joining method suggested three evolutionary groups for these mitogenomes, closely related to A. m. jemenitica, A. m. lamarckii, A. m. syriaca, and A. m. scutellata.

Reproductive biology and morphology of Apis mellifera jemenitica (Apidae) queens and drones.

The current study aimed to investigate the important reproductive biology and morphology of A.m. jemenitica queens and drones through measuring the weight of virgin and mated queens, size and weight of spermathecae, weight of ovaries, number of ovarioles, quantity and viability of semen in queen and drones. Accordingly, the average weights of 0.139 ±â€¯0.01 g and 0.143 ±â€¯0.013 g recorded for virgin and mated queens respectively. The sizes of spermathecae were 1.248 ±â€¯0.103 mm and 1.25 ±â€¯0.022 mm for virgin and mated queens respectively. The mean weight of ovaries was 0.013 ±â€¯0.003 g and the numbers of ovarioles varied from 124 to 163 with the mean of 142.9 ±â€¯9.47 and with no significant difference between virgin and mated queens. The average number of stored sperm per spermathecae of mated queen was estimated to be 4.202 ±â€¯0.613 million with the viability of 80.39%. The average number of sperm per drone recorded was 8,763,950 ±â€¯1,633,203.15 with viability of 79.54 ±â€¯6.70%. In general, the current study revealed that the values recorded for reproductive biology and morphological characters of A. m. jemenitica queens and drones were relatively lower than values recorded for other Apis mellifera races. This mainly could be associated with the body size of the race which is known to be the smallest race among A. mellifera races. Moreover, the harsh environmental conditions of the regions, high temperature, low humidity and limited resources may have contributed for the smaller biological and morphological values. The information will serve as a base in future selection and breeding of program of the race.

Survey and molecular detection of Melissococcus plutonius, the causative agent of European Foulbrood in honeybees in Saudi Arabia.

A large-scale field survey was conducted to screen major Saudi Arabian beekeeping locations for infection by Melissococcus plutonius. M. plutonius is one of the major bacterial pathogens of honeybee broods and is the causative agent of European Foulbrood disease (EFB). Larvae from samples suspected of infection were collected from different apiaries and homogenized in phosphate buffered saline (PBS). Bacteria were isolated on MYPGP agar medium. Two bacterial isolates, ksuMP7 and ksuMP9 (16S rRNA GenBank accession numbers, KX417565 and KX417566, respectively), were subjected to molecular identification using M. plutonius -specific primers, a BLAST sequence analysis revealed that the two isolates were M. plutonius with more than 98% sequence identity. The molecular detection of M. plutonius from honeybee is the first recorded incidence of this pathogen in Saudi Arabia. This study emphasizes the need for official authorities to take immediate steps toward treating and limiting the spread of this disease throughout the country.

Geographical distribution and molecular detection of Nosema ceranae from indigenous honey bees of Saudi Arabia.

The aim of the study was to detect the infection level of honey bees with Nosema apis and/or Nosema ceranae using microscopic and molecular analysis from indigenous honeybee race of eight Saudi Arabian geographical regions. A detailed survey was conducted and fifty apiaries were chosen at random from these locations. Infection level was determined both by microscope and Multiplex-PCR and data were analyzed using bioinformatics tools and phylogenetic analysis. Result showed that N. ceranae was the only species infecting indigenous honeybee colonies in Saudi Arabia. As determined by microscope, Nosema spores were found to be in 20.59% of total samples colonies, while 58% of the samples evaluated by PCR were found to be positive for N. ceranae, with the highest prevalence in Al-Bahah, a tropical wet and dry climatic region, whereas low prevalence was found in the regions with hot arid climate. Honeybees from all eight locations surveyed were positive for N. ceranae. This is the first report about the N. ceranae detection, contamination level and distribution pattern in Saudi Arabia.

Fertility and reproductive rate of Varroa mite, Varroa destructor, in native and exotic honeybee, Apis mellifera L., colonies under Saudi Arabia conditions.

Varroa mite is the most destructive pest to bee colonies worldwide. In Saudi Arabia, preliminary data indicated high infestation levels in the exotic honeybee colonies; such as Apis mellifera carnica and Apis mellifera ligustica, compared to native honeybee subspecies Apis mellifera jemenitica, which may imply higher tolerance to Varroasis. In this study, fertility and reproductive rate of Varroa mite, Varroa destructor, in capped brood cells of the native honeybee subspecies were investigated and compared with an exotic honeybee subspecies, A. m. carnica. Mite fertility was almost alike (87.5% and 89.4%) in the native and craniolan colonies respectively. Similarly, results did not show significant differences in reproduction rate between both subspecies (F = 0.66, Pr > F = 0.42). Number of adult Varroa daughters per fertile mother mite was 2.0 and 2.1 for native and craniolan honeybee subspecies respectively. This may indicate that mechanisms of keeping low infestation rates in the native honeybee colonies are not associated with Varroa reproduction. Therefore, potential factors of keeping lower Varroa infestation rates in native honey bee subspecies should be further investigated.

Chemical compositions and characteristics of organic compounds in propolis from Yemen.

Propolis is a gummy material made by honeybees for protecting their hives from bacteria and fungi. The main objective of this study is to determine the chemical compositions and concentrations of organic compounds in the extractable organic matter (EOM) of propolis samples collected from four different regions in Yemen. The propolis samples were extracted with a mixture of dichloromethane and methanol and analyzed by gas chromatography-mass spectrometry (GC-MS). The results showed that the total extract yields ranged from 34% to 67% (mean = 55.5 ± 12.4%). The major compounds were triterpenoids (254 ± 188 mg g-1, mainly α-, ß-amyryl and dammaradienyl acetates), n-alkenes (145 ± 89 mg g-1), n-alkanes (65 ± 29 mg g-1), n-alkanoic acids (40 ± 26 mg g-1), long chain wax esters (38 ± 25 mg g-1), n-alkanols (8 ± 3 mg g-1) and methyl n-alkanoates (6 ± 4 mg g-1). The variation in the propolis chemical compositions is apparently related to the different plant sources. The compounds of these propolis samples indicate that they are potential sources of natural bio-active compounds for biological and pharmacological applications.