Molecular Variation Between RT-PCR Detected Rotavirus Infection of Naturally Diarrheic Neonatal Calves and Rotavirus Strains of Commercial Vaccines

Neonatal diarrhea is the main cause of morbidity and mortality in calves, and Rotavirus is the main viral etiology. Rotavirus vaccines are one of the main important methods for control of diarrhea in neonates' calves. In the current study, Deoxyribonucleic acid (DNA) sequencing and phylogenetic analysis of Bovine Rotavirus Group A (BRVA) were performed in our study. 1 Calf guard® vaccine genotype (G6P1) and 5 different field genotypes (2 G6P5, 1 G10P5, G10P? and 1 G10P11) were subjected to DNA sequencing. We observed that at the nucleotide level, G10P5 and G10P? sequences were 100 % identical with each other, two G6P5 sequences were 100% identical with each other and there was no significant similarity between sequences of G10P11 with sequences of G6P5, G10P5, and G10P? The phylogenetic analysis of G10P5 and G10P? isolates showed a close cluster with G10 isolates of Sharkia governorate, Egypt, phylogenetic analysis of two G6P5 and one G10P11 isolate showed a close cluster with the VP4 gene of Rotavirus isolates of Dakahlia governorate, Egypt. Molecular comparison between detected and typed Rotaviruses' genotypes with other genotypes of common vaccines indicated that there were genetically close or distance between field and vaccine Rotavirus strains.

Bridging the Species Gap: Morphological and Molecular Comparison of Feline and Human Intestinal Carcinomas

Limited availability of in vivo experimental models for invasive colorectal cancer (CRC) including metastasis and high tumor budding activity is a major problem in colorectal cancer research. In order to compare feline and human intestinal carcinomas, tumors of 49 cats were histologically subtyped, graded and further characterized according to the human WHO classification. Subsequently, feline tumors were compared to a cohort of 1004 human CRC cases. Feline intestinal tumors closely resembled the human phenotype on a histomorphological level. In both species, adenocarcinoma not otherwise specified (ANOS) was the most common WHO subtype. In cats, the second most common subtype of the colon (36.4%), serrated adenocarcinoma (SAC), was overrepresented compared to human CRC (8.7%). Mucinous adenocarcinoma (MAC) was the second most common subtype of the small intestine (12.5%). Intriguingly, feline carcinomas, particularly small intestinal, were generally of high tumor budding (Bd) status (Bd3), which is designated an independent prognostic key factor in human CRC. We also investigated the relevance of feline CTNNB1 exon 2 alterations by Sanger sequencing. In four cases of feline colonic malignancies (3 ANOS, 1 SAC), somatic missense mutations of feline CTNNB1 (p.D32G, p.D32N, p.G34R, and p.S37F) were detected, indicating that mutational alterations of the WNT/β-catenin signaling pathway potentially play an essential role in feline intestinal tumorigenesis comparable to humans and dogs. These results indicate that spontaneous intestinal tumors of cats constitute a useful but so far underutilized model for human CRC. Our study provides a solid foundation for advanced comparative oncology studies and emphasizes the need for further (molecular) characterization of feline intestinal carcinomas.

1164. Acquisition of Methicillin-Susceptible Staphylococcus aureus in the Neonatal Intensive Care Unit: Molecular Comparison of Relatedness of Isolates

Background In the NICU MSSA infections occur frequently and cause morbidity and mortality. Colonization is a risk factor for infection. Optimal infection prevention strategies await a more complete understanding of acquisition and transmission. To investigate possible transmission, we studied whether newly MSSA colonized infants share a strain with another contemporaneously colonized infant. Methods This is a prospective observational study in a level IV NICU from April through November 2019. Infants had weekly MSSA nasal surveillance cultures. Isolates from newly MSSA colonized infants and other infants colonized with MSSA during the same/previous week were subjected to staphylococcal protein A (spa) typing; most pairs with a concordant (CC) spa type were analyzed by whole genome sequencing (WGS; Illumina MiSeq). Pairs of isolates with a CC spa type and < 25 single nucleotide polymorphism differences on WGS were considered closely related (CC pairs). A control group consisted of pairs of isolates from a newly colonized infant with one randomly chosen colonized infant with a discordant (DC) spa type during the same/previous week. The medical records were reviewed for staff member (SM) and room assignment. Fischer’s exact test was used to compare proportions. Results Isolates from 60/68 consecutive newly MSSA colonized infants and 111/133 comparison infants were available for spa typing. Of these 60 infants, 23 (38 %) had a CC spa type with another infant colonized during the same/previous week. Of 18 isolate pairs from infants with a CC spa type that were subjected to WGS, 12 (67%) pairs of isolates were closely related. 7/12 (58 %) of CC pairs had a SM in common compared to 2/13 (15 %) in the DC pair groups, p=0.04. 2/12 (17 %) of CC pairs shared a room compared to 2/13 (15 %) pairs in the DC group, p=1.0. Conclusion Among newly MSSA colonized infants at least 25% are colonized with an isolate closely related to that of another colonized infant indicating likely infant to infant transmission. WGS is more discriminatory than spa typing for MSSA. Given the lack of commonality of room assignment and the commonality of SM assignment, a possible role of healthcare personnel in MSSA transmission should be further investigated. Disclosures Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)

Morphological description of a new specimen of Herpetoreas burbrinki Guo et al 2014 (Serpentes: Colubridae)

The original description of Burbrink’s Keelback, Herpetoreas burbrinki was based on a sole damaged specimen collected from Zayu County, Xizang Autonomous Region, China in September 2007. On 16 August 2019, we collected a second live adult female specimen from the type locality. The identity of the species is established based on morphological and molecular comparison with the holotype. One mitochondrial gene (Cytb) and three nuclear genes (C-mos, Rag1, NT3) of the new specimen were sequenced. The four sequences all share the same haplotypes with the holotype. We describe the coloration in life, variation with the type and expand the morphological description of this species.

Molecular Comparison of Screening Technique for Hepatitis B Virus Infection among Blood Donors in Ibadan, South West, Nigeria

Hepatitis B infection has been a great threat to transfusion medicine and public medicine, especially Nigeria where approximately 18 million Nigerians are chronic carriers. The blood donors in Ibadan are routinely screened with rapid technique or enzyme-linked immunosorbent assay (ELISA) for the detection of Hepatitis B surface antigen (HBsAg), there is paucity of information on the use of Nested Polymerase Chain Reaction (Nested PCR) for the detection of Hepatitis B virus deoxyribonucleic acid (DNA) for the screening of blood donors. This study was aimed at carrying out molecular comparison of the screening techniques for the detection of HBV infection among blood donors in Ibadan, South West, Nigeria. Blood samples were collected from 150 potential blood donors at the blood bank, University College Hospital, Ibadan. Rapid immune-chromatographic technique and nested PCR using primer SF, 979 and MF specific for DNA polymerase genome was used to screen the serum of the blood donors. Processing and analysis of data were performed using IBM statistical package for social sciences (IBM SPSS version 21.0 computer software). Descriptive statistics were presented using chart and tables while statistical significance was taken as P<0.05 The Rapid Screening test showed that 16 (10.7%) of the blood donors were positive while 134 (89.3%) were negative. The molecular detection of the Hepatitis B virus-DNA using nested PCR showed that 7 (4.7%) of the blood donors were positive while 143 (95.3%) were negative. It was also observed that 5 (71.4%) out of the 16 donors (10.7%) captured by the rapid screening were also detected by the Nested PCR, while the remaining 2 (28.6%) detected by the PCR were negative with the Rapid Screening test. The age range of 30 to 39 years and 40 to 49 years had the higher rate of infection 42.9% respectively. Result of the effects of different risk factors generated with the aid of questionnaire reflected that multiple sex partner have the highest prevalence of 16.7% compared to other risk factors In conclusion, the detection of HBV-DNA using nested PCR among blood donors that was positive in Ibadan South West, Nigeria has public health implication for prevention of Hepatitis B virus and this confirms the practice of improper screening of blood before transfusion. Nested PCR techniques helps in early detection of hepatitis B virus DNA among blood donors, due to its high specificity and sensitivity than Rapid technique hence it serves as a confirmatory technique.

Producing a superior genotype from agria potato cultivar using somaclonal variation

This study was performed to produce a superior genotype from Agria potato cultivar using somaclonal variation. Two leaf and meristem explants in combination with four doses of 2,4-D (0, 2, 3 and 4 mg / l) were used for callus induction in a factorial model based on a completely randomized design with 3 replications. Results showed that the meristem explant, along with 3 mg 2,4-D produced the most suitable calluses. In the mentioned regeneration media, the best calluses were regenerated and one of the regenerated genotypes, which was very different from the parent cultivar was selected. The regenerated genotype, was compared with the maternal genotype (Agria) and a control cultivar (Sante), in a field experiment based on a randomized complete block design with 3 replications. The results showed that in terms of most of the studied traits such as tuber weight per plant, stolon length, percentage of dry matter and percentage of starch, the new genotype was superior, compared to the parent cultivar and in terms of peel percentage and maturity date, the parental cultivar was superior. The results of the molecular comparison also showed that based on CBDP marker, both in terms of band number and band size, there were differences between the new genotype and the parental cultivar. In general, results showed that somaclonal variation can be an effective method to generate new genotypes with superior characteristics.

Evolution of the vertebrate head is revealed by the rosette mesoderm

The vertebrate head comprises characteristic combinations of the cranium, brain, cranial nerves and head muscles. However, there have long been arguments about the developmental and evolutionary origins and the possible segmental nature of the head muscles, particularly those anterior to the otic vesicle. In gnathostomes, the presence of pre-otic segments (trunk somite homologs) has been denied by anti-segmentalists, but championed by segmentalists, who have focused on marginally detectable somitomeres or on more obvious head cavities. This metameric ideology has generated various definitions of segments, causing great controversy1,2, and the evaluation of such arguments has been impeded by the relative paucity of relevant work on the head mesoderm of cyclostomes (hagfishes and lampreys). Here, we demonstrate the presence of rosettes (which are reminiscent of somites) in the head mesoderm of lamprey (Lethenteron camtschaticum) embryos using confocal laser scanning and transmission electron microscopy. These transient rosettes, which were not segmented by acellular fissures and were genetically distinct from trunk somites, emerged several times during character individualization of the head muscles. This specialty of the rosette dynamics suggests that the lamprey head mesoderm evolved de novo, rather than from somites, and molecular comparison among deuterostomes supported this perspective.

Molecular Typing of Listeria monocytogenes IVb Serogroup Isolated from Food and Food Production Environments in Poland

Listeria monocytogenes is one of the most important foodborne pathogens that may be present in food and in food processing environments. In the present study, 91 L. monocytogenes isolates of serogroup IVb from raw meat, ready-to-eat food and food production environments in Poland were characterized by whole genome sequencing (WGS). The strains were also compared, using core genome multi-locus sequence typing (cgMLST) analysis, with 186 genomes of L. monocytogenes recovered worldwide from food, environments, and from humans with listeriosis. The L. monocytogenes examined belonged to three MLST clonal complexes: CC1 (10; 11.0% isolates), CC2 (70; 76.9%), and CC6 (11; 12.1%). CC1 comprised of two STs (ST1 and ST515) which could be divided into five cgMLST, CC2 covered two STs (ST2 and ST145) with a total of 20 cgMLST types, whereas CC6 consisted of only one ST (ST6) classified as one cgMLST. WGS sequences of the tested strains revealed that they had several pathogenic markers making them potentially hazardous for public health. Molecular comparison of L. monocytogenes strains tested in the present study with those isolated from food and human listeriosis showed a relationship between the isolates from Poland, but not from other countries.