A Molecular Survey of Bacterial Species in the Guts of Black Soldier Fly Larvae (Hermetia illucens) Reared on Two Urban Organic Waste Streams in Kenya

Globally, the expansion of livestock and fisheries production is severely constrained due to the increasing costs and ecological footprint of feed constituents. The utilization of black soldier fly (BSF) as an alternative protein ingredient to fishmeal and soybean in animal feed has been widely documented. The black soldier fly larvae (BSFL) used are known to voraciously feed and grow in contaminated organic wastes. Thus, several concerns about their safety for inclusion into animal feed remain largely unaddressed. This study evaluated both culture-dependent sequence-based and 16S rDNA amplification analysis to isolate and identify bacterial species associated with BSFL fed on chicken manure (CM) and kitchen waste (KW). The bacteria species from the CM and KW were also isolated and investigated. Results from the culture-dependent isolation strategies revealed that Providencia sp. was the most dominant bacterial species detected from the guts of BSFL reared on CM and KW. Morganella sp. and Brevibacterium sp. were detected in CM, while Staphylococcus sp. and Bordetella sp. were specific to KW. However, metagenomic studies showed that Providencia and Bordetella were the dominant genera observed in BSFL gut and processed waste substrates. Pseudomonas and Comamonas were recorded in the raw waste substrates. The diversity of bacterial genera recorded from the fresh rearing substrates was significantly higher compared to the diversity observed in the gut of the BSFL and BSF frass (leftovers of the rearing substrates). These findings demonstrate that the presence and abundance of microbiota in BSFL and their associated waste vary considerably. However, the presence of clinically pathogenic strains of bacteria in the gut of BSFL fed both substrates highlight the biosafety risk of potential vertical transmission that might occur, if appropriate pre-and-postharvest measures are not enforced.

Trypanosoma rangeli Genetic, Mammalian Hosts, and Geographical Diversity from Five Brazilian Biomes

Trypanosoma rangeli is a generalist hemoflagellate that infects mammals and is transmitted by triatomines around Latin America. Due to its high genetic diversity, it can be classified into two to five lineages. In Brazil, its distribution outside the Amazon region is virtually unknown, and knowledge on the ecology of its lineages and on host species diversity requires further investigation. Here, we analyzed 57 T. rangeli samples obtained from hemocultures and blood clots of 1392 mammals captured in different Brazilian biomes. The samples were subjected to small subunit (SSU) rDNA amplification and sequencing to confirm T. rangeli infection. Phylogenetic inferences and haplotype networks were reconstructed to classify T. rangeli lineages and to infer the genetic diversity of the samples. The results obtained in our study highlighted both the mammalian host range and distribution of T. rangeli in Brazil: infection was observed in five new species (Procyon cancrivorous, Priodontes maximum, Alouatta belzebul, Sapajus libidinosus, and Trinomys dimidiatus), and transmission was observed in the Caatinga biome. The coati (Nasua nasua) and capuchin monkey (S. libidinosus) are the key hosts of T. rangeli. We identified all four T. rangeli lineages previously reported in Brazil (A, B, D, and E) and possibly two new genotypes.

MORPHOLOGICAL AND GENOMIC VARIABLITY OF SCLEROTIUM ROLFSII SACC. CAUSING STEM ROT OF TUBEROSE

Tuberose is one of the most important ornamental bulbous flower crop cultivated for cut and loose flower trade. The flower has been used for ornaments, bouquets and buttonholes or crown and frequently used during marriages and religious ceremonies. The tuberose was often infected by various numbers of diseases; among that S. rolfsii is the one of the major disease which causes stem rot disease. The S. rolfsii were collected from various locations of Tamil Nadu were examined for morphological and genomic variability. Fifteen isolates of S. rolfsii was assessed and various morphological growth parameters (Mycelia growth, No. of sclerotia/ plate, colour of sclerotia, time taken for sclerotial production (days) and variations among S. rolfsii isolates was recorded. ITS region of rDNA amplification with specific ITS1 and ITS4 universal primers produced approximately 600 to 700 bp in all the isolates confirmed that all the isolates obtained are S. rolfsii. The sequences of isolates viz., Sr1 and Sr2 were identified as S. rolfsii through BLAST search in NCBI website (www.blast.ncbi.nlm.nih.gov/Blast). The sequences were deposited in the Gene Bank with the accession numbers MK880692, MK880693.

Efficient Ammonium Removal by Bacteria Rhodopseudomonas Isolated from Natural Landscape Water: China Case Study

In this study, we isolated a strain of photosynthetic bacteria from landscape water located in Southwest University, Chongqing, China, and named it Smobiisys501. Smobiisys501 was Rhodopseudomonas sp. according to its cell morphological properties and absorption spectrum analysis of living cells. The analysis of the 16S rDNA amplification sequence with specific primers of photosynthetic bacteria showed that the homology between Smobiisys501 and Rhodopseudomonas sp. was 100%, and the alignment results of protein sequences of the bacterial chlorophyll Y subunit showed that Smobiisys501 and Rhodopseudomonas palustris were the most similar, with a similarity of more than 92%. However, Smobiisys501 could not utilize glucose and mannitol as a carbon source and had a low fatty acid content, which were different from the related strains of the genus Rhodopseudomonas. Moreover, the DNA-DNA relatedness was only 42.2 ± 3.3% between Smobiisys501 and the closest strain Rhodopseudomonas palustris. Smobiisys501 grew optimally at 30 °C and pH 7.0 in the presence of yeast extract, and it could efficiently remove ammonium (99.67% removal efficiency) from synthetic ammonium wastewater. All the results indicated that Smobiisys501 was a novel species of Rhodopseudomonas, with the ability to remove ammonium.

Review of 16S and ITS Direct Sequencing Results for Clinical Specimens Submitted to a Reference Laboratory

We evaluated the performance of 16S and internal transcribed spacer (ITS) region amplification and sequencing of rDNA from clinical specimens, for the respective detection and identification of bacterial and fungal pathogens. Direct rDNA amplification of 16S and ITS targets from clinical samples was performed over a 4-year period and reviewed. All specimens were from sterile sites and submitted to a reference laboratory for evaluation. Results of 16S and ITS were compared to histopathology, Gram and/or calcofluor stain microscopy results. A total of 277 16S tests were performed, with 64 (23%) positive for the presence of bacterial DNA. Identification of an organism was more likely in microscopy positive 16S samples 14/21 (67%), compared to 35/175 (20%) of microscopy negative samples. A total of 110 ITS tests were performed, with 14 (13%) positive. The yield of microscopy positive ITS samples, 9/44 (21%), was higher than microscopy negative samples 3/50 (6%). Given these findings, 16S and ITS are valuable options for culture negative specimens from sterile sites, particularly in the setting of positive microscopy findings. Where microscopy results are negative, the limited sensitivity of 16S and ITS in detecting and identifying an infectious agent needs to be considered.