A Report on Molecular Detection and Phylogenetic Evaluation of Anaplasma marginale in Ticks and Blood Samples Collected from Cattle in District Layyah in Punjab (Pakistan)

Anaplasmosis is a tick-borne disease caused by an obligate gram-negative bacteria, Anaplasma (A.) marginale. This study reports on the seasonal prevalence, epidemiology and phylogeny of A. marginale in three breeds of cattle from Layyah district, Southern Punjab, Pakistan. A total of 844 blood samples (Cross = 300, Holstein Friesian = 244, Sahiwal breed = 300) of apparently healthy cattle on a seasonal basis along with the epidemiological data collected during May 2018 to April 2019. The polymerase chain reaction resulting 265 base pair amplicons specific to a major surface 1b protein-encoding genes of A. marginale in 8.6% (73/844) of cattle registered.

The highest prevalence was observed during the fall (18.3%) followed by summer (9.7%) and winter (7.1%). Holstein Friesian breed is most susceptible to infection A. marginale (13.1%) followed by Sahiwal (7.6%) and cross-breeding (6%). partial gene sequences amplified representation of A. marginale submitted to GenBank (Accession number MK032842 and MK032843). 37/844 (4.3%) Giemsa stained blood smears found positive for Anaplasma spp. a small number of ticks including Hyalomma anatolicum, Hyalomma excavatum, Cattle Tick, Haemaphysalis punctata identified from cattle but none of them were found positive PCR for the presence of A. marginale. Epidemiological factor analysis revealed that women with cows and farm water supplies from the pool, other dairy farms where animals and dogs live with cattle and dogs have fleas burden on them had a significant association with the prevalence of A. marginale.

It was observed that the white blood cells, lymphocytes (%), monocytes (%) hematocrit, mean corpuscular hemoglobin and hemoglobin concentration of living cells on average were significantly impaired in A. marginale-positive than negative cows.

Taxonomy and Molecular phylogenetics of Ensiculiferaceae, fam. November (Peridiniales, Dinophyceae), with consideration of their life history

In the current limitation, Thoracosphaeraceae comprises all dinophytes showed calcification coccoid cells produced during the life of their history. Species hitherto assigned to Ensiculifera and Pentapharsodinium largely based on monadoid life-history stages, while links to coccoid stage (sometimes treated taxonomically different) are not always resolved. We investigate the different stages of life-history and the DNA sequence of the data Ensiculifera mexicana and other species occur in samples collected from around the world.

Based on concatenated ribosomal RNA gene sequences represented Ensiculiferaceae peridinalean different branches, which shows the relationship much to dinophytes other calcareous. Both morphological and molecular data (particularly from coccoid stage) revealed the presence of three distinct clades within Ensiculiferaceae, which may include other dinophytes showing the life-history stages of the parasite. At higher taxonomic levels, Ensiculiferaceae shows the relationship of parasites and endosymbionts (eg, Blastodinium and zooxanthella) and dinophytes save diatom not chloroplasts. These unexpected phylogenetic relationships strengthened by the presence of five plates in all taxa Cingular, which is different from the six plates cingular from most other Thoracosphaeraceae. We here describe Ensiculiferaceae, correct description and Pentapharsodinium Ensiculifera, upright Matsuokaea and give some new combinations at the species level.

Molecule Detection of Potato Virus in Bangladesh and their phylogenetic analysis

Potato (Solanum tuberosum) is a major food source all over the world including Bangladesh. viral diseases is a major obstacle to sustainable potato production by reducing the quality and quantity. To determine the current status of the eight important potato virus in Bangladesh, tuber samples were collected from three main potato-growing region (Munshiganj, Jessore and Bogra district) in January-February 2017 and February 2018.

The reverse transcription polymerase chain reaction (RT-PCR) and protein coat (CP) -specific primer used to amplify the CP sequence of each virus, and confirmed by sequencing, which is stored in GenBank. The results showed that the samples of tubers were subjected to virus Potato leafroll (PLRV), potato virus X (PVX), potato virus Y (PVY), virus Potato S (PVS), virus Potato H (PVH), Potato Aucuba mosaic virus (PAMV) and potato virus M (PVM) infection, whereas the mixed infections are very common. Phylogenetic analysis revealed that PLRV from this study is closely related to Canadian and Chinese isolates, respectively; PVX is closely associated with the Canadian and Chinese isolates, respectively; PVY isolates are closely related to China;

hsa-let-7e Primers
MPH01010	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7f Primers
MPH01011	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7g Primers
MPH01012	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7i Primers
MPH01013	ABM	150 ul / 10 uM	EUR 145.2

rno-let-7b Primers
MP-r00034	ABM	150 ul / 10 uM	EUR 211.2

mmu-let-7b Primers
MPM00008	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7a-3p Primers
MPH02041	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7a-5p Primers
MPH02042	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7c-3p Primers
MPH02045	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7c-5p Primers
MPH02046	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7d-3p Primers
MPH02047	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7d-5p Primers
MPH02048	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7e-3p Primers
MPH02049	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7e-5p Primers
MPH02050	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7f-5p Primers
MPH02053	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7g-3p Primers
MPH02054	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7g-5p Primers
MPH02055	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7i-3p Primers
MPH02056	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7i-5p Primers
MPH02057	ABM	150 ul / 10 uM	EUR 145.2

rno-let-7b-3p Primers
MPR00308	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7b-5p Primers
MPR00309	ABM	150 ul / 10 uM	EUR 211.2

mmu-let-7b-3p Primers
MPM00737	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7b-5p Primers
MPM00738	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7f-1-3p Primers
MPH02051	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7f-2-3p Primers
MPH02052	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7a-2-3p Primers
MPH03980	ABM	150 ul / 10 uM	EUR 145.2

hsa-let-7b RT-PCR Primer Set
abx096770-1nmol	Abbexa	1 nmol	EUR 343.2

rno-let-7d Primers
MP-r00007	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7a Primers
MP-r00033	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7c Primers
MP-r00035	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7e Primers
MP-r00036	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7f Primers
MP-r00037	ABM	150 ul / 10 uM	EUR 211.2

rno-let-7i Primers
MP-r00038	ABM	150 ul / 10 uM	EUR 211.2

mmu-let-7a Primers
MPM00007	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7c Primers
MPM00009	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7d Primers
MPM00010	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7e Primers
MPM00011	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7f Primers
MPM00012	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7g Primers
MPM00013	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7i Primers
MPM00014	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7j Primers
MPM00753	ABM	150 ul / 10 uM	EUR 145.2

mmu-let-7k Primers
MPM00754	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-1 Primers
MPH01014	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-7 Primers
MPH01976	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-9 Primers
MPH02015	ABM	150 ul / 10 uM	EUR 145.2

rno-miR-7b Primers
MP-r00039	ABM	150 ul / 10 uM	EUR 211.2

mmu-miR-7b Primers
MPM00702	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-16 Primers
MPH01191	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-17 Primers
MPH01192	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-21 Primers
MPH01259	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-22 Primers
MPH01277	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-24 Primers
MPH01294	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-25 Primers
MPH01295	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-31 Primers
MPH01329	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-32 Primers
MPH01417	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-93 Primers
MPH02022	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-95 Primers
MPH02035	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-96 Primers
MPH02036	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-98 Primers
MPH02037	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-100 Primers
MPH01015	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-101 Primers
MPH01016	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-103 Primers
MPH01017	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-105 Primers
MPH01019	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-107 Primers
MPH01022	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-10a Primers
MPH01023	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-10b Primers
MPH01024	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-122 Primers
MPH01045	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-124 Primers
MPH01060	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-126 Primers
MPH01082	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-128 Primers
MPH01110	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-132 Primers
MPH01144	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-134 Primers
MPH01151	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-136 Primers
MPH01154	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-137 Primers
MPH01155	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-138 Primers
MPH01156	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-141 Primers
MPH01161	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-143 Primers
MPH01164	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-144 Primers
MPH01165	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-145 Primers
MPH01166	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-147 Primers
MPH01172	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-149 Primers
MPH01178	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-150 Primers
MPH01179	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-152 Primers
MPH01182	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-153 Primers
MPH01183	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-154 Primers
MPH01187	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-155 Primers
MPH01188	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-15a Primers
MPH01189	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-15b Primers
MPH01190	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-182 Primers
MPH01197	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-183 Primers
MPH01201	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-184 Primers
MPH01202	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-185 Primers
MPH01203	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-186 Primers
MPH01204	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-187 Primers
MPH01205	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-18a Primers
MPH01208	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-18b Primers
MPH01209	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-190 Primers
MPH01210	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-191 Primers
MPH01214	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-192 Primers
MPH01221	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-194 Primers
MPH01225	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-195 Primers
MPH01226	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-197 Primers
MPH01229	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-198 Primers
MPH01238	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-19a Primers
MPH01242	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-19b Primers
MPH01243	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-202 Primers
MPH01247	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-203 Primers
MPH01248	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-204 Primers
MPH01249	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-205 Primers
MPH01250	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-206 Primers
MPH01254	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-20a Primers
MPH01257	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-20b Primers
MPH01258	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-210 Primers
MPH01260	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-211 Primers
MPH01261	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-212 Primers
MPH01268	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-214 Primers
MPH01269	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-215 Primers
MPH01270	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-218 Primers
MPH01273	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-221 Primers
MPH01281	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-222 Primers
MPH01282	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-223 Primers
MPH01283	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-224 Primers
MPH01284	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-23a Primers
MPH01291	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-23b Primers
MPH01292	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-23c Primers
MPH01293	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-26a Primers
MPH01296	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-26b Primers
MPH01297	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-27a Primers
MPH01298	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-27b Primers
MPH01299	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-297 Primers
MPH01306	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-298 Primers
MPH01307	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-29a Primers
MPH01310	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-29b Primers
MPH01311	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-29c Primers
MPH01312	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-300 Primers
MPH01313	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-30a Primers
MPH01324	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-30b Primers
MPH01325	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-30c Primers
MPH01326	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-30d Primers
MPH01327	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-30e Primers
MPH01328	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-325 Primers
MPH01431	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-326 Primers
MPH01432	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-328 Primers
MPH01433	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-329 Primers
MPH01434	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-335 Primers
MPH01439	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-33a Primers
MPH01446	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-33b Primers
MPH01447	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-340 Primers
MPH01448	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-345 Primers
MPH01451	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-346 Primers
MPH01452	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-34a Primers
MPH01453	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-34b Primers
MPH01454	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-363 Primers
MPH01486	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-365 Primers
MPH01492	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-367 Primers
MPH01514	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-370 Primers
MPH01545	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-372 Primers
MPH01550	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-373 Primers
MPH01551	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-375 Primers
MPH01555	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-377 Primers
MPH01559	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-378 Primers
MPH01560	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-379 Primers
MPH01563	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-380 Primers
MPH01564	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-381 Primers
MPH01565	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-382 Primers
MPH01566	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-383 Primers
MPH01567	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-384 Primers
MPH01568	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-410 Primers
MPH01606	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-411 Primers
MPH01607	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-412 Primers
MPH01608	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-421 Primers
MPH01609	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-424 Primers
MPH01613	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-425 Primers
MPH01614	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-429 Primers
MPH01654	ABM	150 ul / 10 uM	EUR 145.2

hsa-miR-431 Primers
MPH01675	ABM	150 ul / 10 uM	EUR 145.2

PVS is closely related to China and isolate Iran, respectively; PAMV closely related isolates of Canada; PVH is closely related to isolates Huhhot China; and PVM is closely related to India and isolate Iran, respectively. As far as we know, PAMV in this study is the first report in Bangladesh. These findings will provide great scope for the right virus control strategies for the production of virus-free potato in Bangladesh.